1. Background

Matrix Gel is a soluble basement membrane preparation extracted from EHS mouse tumors rich in extracellular matrix (ECM) proteins. Its main components include laminin, type IV collagen, heparan sulfate proteoglycans (HSPG), entactin/nidogen, as well as growth factors such as TGF-beta, EGF, IGF, FGF, tissue plasminogen activator, and other growth factors endogenous to the EHS tumor. In the mid-1980s, it was independently discovered and characterized by J. Engelbreth-Holm and Richard Swarm from Denmark [1]. The primary production process involves extracting/washing tumor homogenates with saline to collect soluble proteins, followed by solvent extraction to remove insoluble complexes, and finally dialysis at low temperature to obtain a colorless to pale yellow solution.

Table 1: Composition of Matrix Gel [1]

Matrix Gel provides structural support, tensile strength, and scaffolding for tissues and cells. It also serves as a three-dimensional microenvironment for cell adhesion and motility, a reservoir for growth factors, chemokines, and cytokines, and a source of signaling cues for cell morphogenesis and differentiation. YeaSen Biotechnology has developed and manufactured Ceturegel™ Matrix Gel that is free of LDEV (Lactate Dehydrogenase Elevating Virus), has ultra-low endotoxin levels, and has been tested for mycoplasma contamination to ensure sterility. The product line includes standard concentration, high concentration, and reduced growth factor versions.

2. Applications

One major application of Matrix Gel is in organoid culture. Organoids are 3D cell culture systems derived in vitro from stem cells, progenitor cells, or tumor cells, which self-organize into structures resembling native tissue architecture. In 2021, organoid technology was designated as a key national R&D project under China's 14th Five-Year Plan [2]. Compared to traditional 2D disease models, organoids more closely mimic the in vivo environment and are increasingly important for studying disease development, homeostasis, and pathogenesis. They have broad applications in areas such as cell therapy, drug discovery, genetic engineering, immunology, and tissue regeneration.

Figure 1: Applications of Organoids [3]

3. Reagent Preparation

1. Intestinal digestion solution: 500 μL of 0.5 M EDTA (pH 8.0) + 50 mL DPBS + 1% antibiotics (penicillin-streptomycin) + 100 μL primocin

2. Washing solution: sufficient DPBS + 1% antibiotics

3. Rinse solution: sufficient DPBS + 0.1% BSA + 1% antibiotics + 1 mL primocin

4. Experimental Procedures

4.1 2D Culture

1. Thawing and Storage of Ceturegel™ Matrix Gel

[Note]: Ceturegel™ Matrix Gel is highly temperature-sensitive. Avoid repeated freeze-thaw cycles. All steps involving dispensing or preparation prior to gelation must be performed on ice (4 °C), as slight temperature increases may cause premature gelation, leading to uneven distribution or impaired gel formation. All tubes and pipette tips used must be pre-chilled.

a) Upon receipt, if not used immediately, store the entire vial directly at –20 °C (do not use frost-free freezers).

b) For first use, place the entire vial on ice and transfer to 4 °C overnight for complete thawing. It is recommended to centrifuge at 14,000 × g for 20 min at 4 °C and carefully collect the supernatant.

2. Special Notes on Use of Ceturegel™ Matrix Gel

Matrix Gel rapidly gels at 22–35 °C. To ensure optimal gelation performance and stability, the final dilution concentration may vary by lot. Dilute with serum-free medium and use immediately after dilution.

A. Thin Gel Coating Method

1) After thawing, mix Matrix Gel thoroughly using a pre-chilled pipette tip.

2) Place the culture plate on ice and add Matrix Gel at a concentration of 50 μL/cm² of growth surface area.

3) Incubate at 37 °C for 30 min. The plate is then ready for use.

B. Thick Gel Method

1) After thawing, mix Matrix Gel thoroughly using a pre-chilled pipette tip.

2) Place the culture plate on ice. Mix cells with Matrix Gel and suspend evenly using a pre-chilled pipette tip. Add Matrix Gel at a concentration of 150–200 μL/cm² of growth surface area.

3) Incubate at 37 °C for 30 min, then add cell culture medium. Cells may also grow on top of the thick gel layer.

C. Thin Layer Coating Method

1) After thawing, mix Matrix Gel thoroughly using a pre-chilled pipette tip.

2) Dilute Matrix Gel with serum-free medium to the desired concentration. A gradient test is recommended to determine the optimal coating concentration for specific experiments.

3) Add diluted Matrix Gel to the culture vessel, ensuring complete coverage of the growth surface. Incubate at room temperature for 1 hour.

4) Remove unbound Matrix Gel and rinse gently with serum-free medium. The plate is now ready for use.

[Note]: Coated plates should ideally be used the same day, but may be stored depending on application. After adding culture medium, coated plates can be stored at 37 °C for up to 1 week.

D. Validation Procedure

1) After thawing, dilute Matrix Gel 10-fold with pre-chilled PBS and keep on ice.

2) Add 0.5 mL of diluted Matrix Gel to three wells of a non-tissue-culture-treated 6-well plate, ensuring even coverage of the bottom. Incubate at 37 °C for 30–60 min, then remove excess liquid.

3) Digest 293T cells using standard protocol and adjust cell suspension to 1 × 10⁵ cells/mL.

4) Slowly add 3 mL of cell suspension down the side of each well to avoid disturbing the coated layer. Seed the same number of cells in uncoated control wells.

5) Incubate the 6-well plate overnight (12–18 h) in a 37 °C, 5% CO₂ incubator.

6) Observe cell attachment and take photos. Results are shown below (a: untreated, b: coated with 10× diluted Matrix Gel):

Experimental Results

Figure 2: Support of 2D Cell Culture

Troubleshooting

Q: How to ensure sterility during culture?

A: Add 1% antibiotics and an appropriate amount of gentamicin sulfate to the culture medium.

4.2 Invasion Assay

Figure 3: Migration and Invasion Assay Workflow

1. Matrix Gel Coating for Transwell

Thaw Ceturegel™ Matrix Gel and dilute 1:8 with serum-free medium (final concentration should not exceed 3 mg/mL). Add the diluted Matrix Gel to the upper chamber of a Transwell insert to coat the membrane. Incubate at 37 °C for 30 min to allow gelation. Hydrate the basement membrane before use.

2. Cell Suspension Preparation

① Starve cells for 12–24 h to eliminate serum effects (optional).

② Digest cells, centrifuge to remove medium, wash 1–2 times with PBS, and resuspend in serum-free medium containing BSA. Adjust cell density to 4 × 10⁴ – 2 × 10⁵ cells/mL.

3. Cell Seeding

① Add 100 μL of cell suspension to the upper chamber of the Transwell.

② Add 600 μL of medium containing 20% FBS to the lower chamber. Avoid introducing bubbles between the lower chamber and the insert, as bubbles may weaken or eliminate the chemotactic gradient.

③ Culture cells for 12–48 h (duration depends on the invasive capacity of the cancer cells).

4. Data Analysis

Remove the Transwell insert, discard medium, wash twice with Ca²⁺-free PBS, fix with 4% paraformaldehyde for 30 min, and air-dry briefly.

Stain with 0.1% crystal violet for 20 min, gently wipe off non-migrated cells from the upper surface with a cotton swab, wash three times with PBS, and count cells in five random fields under a 400× microscope.

Experimental Results

Figure 4: Crystal violet staining of cells after invasion assay

Troubleshooting

Q1: What precautions should be taken when handling Matrix Gel?

A: All operations must be performed under sterile conditions. Use pre-chilled pipettes to maintain Matrix Gel in a liquid, homogeneous state.

Q2: How to aliquot and store Matrix Gel?

A: After thawing, aliquot Ceturegel™ Matrix Gel into multiple small tubes using pre-chilled cryovials. Freeze rapidly and store at –20 °C. Avoid repeated freeze-thaw cycles. All items used during handling (pipettes, tips, tubes) must be pre-chilled.

4.3 Angiogenesis Assay

1. Matrix Gel Preparation

1) The night before the experiment, remove Ceturegel™ Matrix Gel from the freezer and thaw overnight at 4 °C. Pre-chill all labware.

2) Keep Matrix Gel on ice throughout the experiment.

3) Open the sterile packaging of the angiogenesis slide and remove the slide.

4) Add 10 μL of Ceturegel™ Matrix Gel into each well. Hold the pipette tip vertically above the center of the well to prevent Matrix Gel from flowing into the upper chamber and leaving residual gel.

2. Gelation

1) Cover the slide with its lid. Prepare a 10 cm culture dish with wetted paper towels to create a humidified chamber.

2) Place the slide into the dish and cover with the dish lid.

3) Incubate the entire dish in a CO₂ incubator at 37 °C for ~30 min to allow gelation. Meanwhile, prepare the cell suspension.

3. Cell Seeding

1) Prepare a cell suspension at a density of 2 × 10⁵ cells/mL after digestion, and mix thoroughly.

2) Remove the slide with gelled Matrix Gel.

3) Add 50 μL of cell suspension to each well, keeping the pipette tip vertical and avoiding contact with the gel below.

4) Add culture medium, cover the slide, and incubate. Cells will settle onto the surface of the Matrix Gel over time.

4. Imaging

Take photos at regular intervals according to cell growth rate.

5. Immunofluorescence Staining (Optional)

1) Carefully remove the medium without disturbing the gel or cell network. Dilute calcein AM in serum-free medium to a final concentration of 6–8 µg/mL.

2) Add staining solution to fully cover the cells. Incubate at room temperature, protected from light, for 30–40 min.

3) Wash three times with PBS, adding PBS gently to the upper chamber to avoid dislodging cells.

4) Image using Ex = 485 nm, Em = 529 nm.

6. Data Analysis

Measure and record tube length, coverage area, number of loops, and number of branch points, followed by statistical analysis.

Experimental Results

Figure 5: Angiogenesis results and immunofluorescence staining images

Troubleshooting

1. All pipette tips, culture plates, and EP tubes that contact Matrix Gel must be pre-chilled.

2. Avoid air bubbles during Matrix Gel coating to prevent interference with imaging.

3. Seed cells evenly to avoid clustering, which may affect uniform tube formation.

4. Handle gently during seeding to avoid damaging the gel surface, which could affect imaging.

5. Maintain appropriate cell suspension temperature; low temperature may melt the gel, impairing cell migration and tubulogenesis.

4.4 In Vivo Tumor Formation

Using a HepG2 cell xenograft tumor model in nude mice as an example, Ceturegel™ Matrix Gel is mixed with cell suspension at a 1:1 ratio and subcutaneously injected into 4–5 week-old female BALB/c-nu mice. The experimental workflow is as follows:

1. Prepare HepG2 cells in logarithmic growth phase with 80–90% confluence. Change to fresh medium the night before cell collection.

2. Digest cells with trypsin. When cells begin to round up but have not detached, remove trypsin and resuspend in serum-free medium. Centrifuge once, then resuspend to a final concentration of 1 × 10⁸ cells/mL.

3. At 4 °C, mix cell suspension and Ceturegel™ High Concentration Matrix Gel at a 1:1 ratio to achieve a final concentration of 5 × 10⁷ cells/mL.

4. Hold the nude mouse with the left hand and inject subcutaneously at the right shoulder. Insert the needle deeper (~1 cm) under the skin to minimize leakage of the cell suspension after injection. Inject 200 μL. (Complete the process within 30 min; keep the cell suspension on ice during the procedure to reduce apoptosis and prevent premature gelation.)

5. Return the mouse to the cage for continued housing. Tumors typically appear within 1–4 weeks. Euthanize the mouse according to experimental design when tumor volume does not exceed 1000 mm³, and take photos.

[Note]: The control group consists of a mixture of medium and cell suspension at the same final cell density as the Matrix Gel group.

Experimental Results

Troubleshooting

1.Can normal mice be used for this experiment?

Nude mice lack a thymus and have compromised immunity, making them unable to reject tumor cells, thus facilitating tumor formation.

2.Why is the tumor not growing?

① Tumor growth may take 1–2 months.

② Possible reasons include insufficient cell number, poor cell viability, or the cell line’s low tumorigenicity, which may require supportive factors such as Matrix Gel.

3. It is recommended to use immunodeficient mice aged 4–6 weeks.

4.5 3D Culture

Cell Suspension Preparation

Pre-chill 24-well plates and pipette tips on ice. Keep Matrix Gel on ice or thaw slowly at 4 °C. Harvest HepG2 cells in logarithmic growth phase, gently rinse with PBS, and digest with trypsin. After digestion, prepare and collect a single-cell suspension. Centrifuge at 1500 rpm for 3 min, discard supernatant, and resuspend in complete DMEM medium. Count cells using a hemocytometer and adjust concentration to 1.5 × 10⁵ cells/mL.

Cell Seeding

Mix thawed Matrix Gel and adjusted single-cell suspension at a 1:1 ratio on ice with gentle mixing. Using a pre-chilled 200 μL pipette tip, transfer 40–50 μL of the mixture and dispense vertically into the center of pre-chilled 24-well plates to form dome-shaped droplets.

Data Analysis

After incubating in a 37 °C, 5% CO₂, humidified incubator for 30 min to stabilize, add 500 μL to 2 mL of complete DMEM medium per well and continue culture. Observe and photograph daily.

Experimental Results

Figure 6. Results of 3D cell culture after 4 days

Troubleshooting

1.All pipette tips, culture plates, and EP tubes contacting Matrix Gel must be pre-chilled.

2.HepG2 cells tend to aggregate; ensure thorough resuspension in medium.

3.Avoid contact between the cell-gel droplet and the edge of the well during seeding.

4.Between days 4–10, 3D cells grow rapidly; replace with fresh medium regularly.

4.6 Organoid Culture

4.6.1 Mouse Intestinal Organoids

1. Sample Preparation: Euthanize mouse by cervical dislocation and disinfect the surface with alcohol. Under sterile conditions, excise 3–15 cm of small intestine from the region near the stomach. Carefully remove mesentery and fat from the outer surface using forceps and place the tissue into pre-chilled DPBS containing 1% antibiotics at 4 °C.

2. Sample Washing: Flush the lumen 2–3 times with a syringe. Open the intestine with scissors, place the luminal side up, and gently scrape off villi using a surgical blade. When villi are completely removed (tissue appears translucent), transfer the tissue to a new dish containing DPBS and wash 2–3 times.

3. Initial Processing: Cut the washed intestinal tissue into ~2 mm fragments and transfer to a new 50 mL conical tube. Wash gently 3–5 times with DPBS to remove villus cells and floating fat.

4. Tissue Digestion: Add 10–15 mL of pre-chilled DPBS containing 3–5 mM EDTA to the intestinal fragments and incubate at 4 °C for ~30 min, gently shaking the tube every 10 min.

5. After digestion, discard the EDTA supernatant and gently rinse the tissue 2–3 times with fresh DPBS to remove residual EDTA.

6. Add 10–15 mL of pre-chilled DPBS containing 0.1% BSA to the intestinal fragments. Pipette repeatedly to dissociate crypts from the basement membrane. Examine a small aliquot under a microscope; stop pipetting when abundant crypt-like structures are observed. Filter the suspension through a 70 μm cell strainer and collect the flow-through.

7. Repeat steps 5–6 twice, then centrifuge at 1500 rpm, 4 °C for 3 min.

8. Mixture Preparation: Resuspend the crypt pellet in Ceturegel™ Matrix Gel. Use approximately 200–600 crypts per 10 μL of Matrix Gel. Keep the suspension on ice and proceed quickly to prevent premature gelation.

[Note]: The Matrix Gel dilution ratio should be ≥50% to ensure structural stability during culture.

9. Seed 30–50 μL of the mixture into the center of each well of a 24-well plate, avoiding contact with the well walls.

10. Place the plate in a 37 °C CO₂ incubator and incubate for ~30 min to allow gelation.

11. After complete gelation of Ceturegel™ Matrix Gel, slowly add pre-prepared intestinal organoid culture medium along the wall of each well (800 μL/well).

12. Culture the 24-well plate in a 37 °C CO₂ incubator. Replace with fresh medium every 3 days and monitor organoid growth. Mouse intestinal organoids typically form within 5–7 days.

Experimental Results 

 Figure 7: In vitro culture results of mouse small intestinal organoids

Problem Analysis

1.What digestion solution should be used for tissue digestion, and what is the digestion duration?

Tissue can be digested using either EDTA or collagenase. For hollow organs such as the small intestine and stomach, EDTA is suitable. Collagenase comes in different types with broad tissue applicability and may be combined with DNase. Digestion times vary significantly across tissues, ranging from several minutes to several hours.

2.Volumes of matrix gel and culture medium required per well for organoid culture in different plate formats?

Commonly used plates for organoid culture include 24-well plates, where a 50 μL droplet of matrix gel is dispensed per well, overlaid with 500 μL of culture medium; 96-well plates use 10 μL of matrix gel per well, covered with 200 μL of medium; for 6-well plates, multiple 50 μL gel droplets can be seeded per well, with 2–3 mL of medium added to cover the droplets.

4.6.2 Tumor Organoids

1. Pre-experiment Preparation:

Thaw tumor organoid culture kit components and matrix gel at 4°C.

Prepare sufficient volumes of DPBS containing 1% penicillin-streptomycin (dual antibiotics) and DPBS containing 0.1% BSA.

Prepare one-fifth strength trypsin solution: mix 32 mL basal medium with 8 mL trypsin, pre-warm at 37°C for later use.

2. Sampling:

Under sterile conditions, excise approximately 2 cm of viable tumor tissue near the gastric end. Use forceps to remove surrounding connective tissues, fat, fascia, etc., and place the tissue into ice-cold DPBS (supplemented with dual antibiotics) or culture medium for transport.

3. Washing:

Using surgical scissors, mince the tumor tissue into ~2 mm fragments. Transfer the minced tissue pieces into a new 50 mL conical tube containing DPBS and wash until the supernatant becomes clear. Discard supernatant and add Type I collagenase (~3× volume of tumor tissue). Digest for 30 min, followed by two washes with basal medium.

4. Transfer the collagenase-digested tumor tissue into basal medium containing one-fifth strength trypsin. Incubate at 37°C for 20–30 min. After digestion, add an appropriate volume of basal medium containing 0.1% BSA to neutralize the reaction. Gently mix and collect the supernatant.

5. Add 30 mL of basal medium containing 0.1% BSA to the remaining pellet. Resuspend and triturate the tissue fragments using a 10 mL pipette in a 50 mL conical tube—repeatedly pass the tissue through the pipette tip to generate mechanical shearing forces that dissociate tumor cells from the basement membrane. Repeat 6–10 times. Take a small aliquot for microscopic examination. Stop trituration when abundant single-cell structures are observed. Filter the suspension through a 70 μm cell strainer.

6. Collect the flow-through fraction from filtration. Centrifuge at 300 × g, 4°C for 3 min.

Discard supernatant. Resuspend the pellet in 1 mL of basal medium containing 0.1% BSA. Take 20 μL for microscopy and cell counting. After counting, transfer the desired number of cells into a new tube, centrifuge again at 300 × g for 3 min, discard supernatant, and keep on ice.

7. Resuspend the cancer cell pellet in an appropriate volume of organoid-specific growth medium. Recommended resuspension density: 1–2 × 10⁶ cells/mL. Keep the tube on ice after resuspension. Resuspension time should not exceed 30 seconds to prevent premature gelation of matrix gel.

[Note]: The final dilution ratio of matrix gel should remain above 50% to ensure structural stability during culture.

8.Deposit the mixture of matrix gel and cell suspension as ~30 μL droplets in the center of each well of a 24-well plate. Avoid contact between droplets and the sidewalls of the wells.

[Note]: To prevent premature solidification at room temperature, this step must be performed quickly.

9.Place the inoculated plate in a 37°C CO₂ incubator and incubate for 3–5 min. Then invert the plate and incubate for another 10–20 min to allow complete polymerization of the matrix gel. Once fully solidified, slowly add pre-warmed complete tumor organoid medium along the side of each well—750 μL per well for 24-well plates. Avoid disrupting the already formed gel structure during medium addition.

10.Culture the 24-well plate in a 37°C CO₂ incubator. Change the medium every 3 days, taking care not to disturb the matrix gel. Monitor organoid growth closely. Under optimal conditions, tumor organoids should form within 5 to 10 days.

Figure 8: Cervical cancer organoids cultured for 7 days

Figure 9: Colon cancer organoids cultured for 6 days

Figure 10: Pancreatic cancer organoids cultured for 9 days

Figure 11: Ovarian cancer organoids cultured for 7 days

Problem Analysis

1. All pipette tips, culture plates, and EP tubes that come into contact with matrix gel must be pre-chilled.

2. Serum must not be added at any stage throughout the entire culture process.

4.7 Hematoxylin and Eosin (H&E) Staining of Organoids

4.8 Sequencing-Based Identification of Organoids

Related Reagents

1. Tissue and Cell Dissociation

Collagenase refers to a class of enzymes capable of specifically recognizing the Pro-X-Gly-Pro sequence (highly prevalent in collagen and rare in other proteins), cleaving the peptide bond between the neutral amino acid (X) and glycine (Gly), and hydrolyzing collagen in connective tissues. The most effective collagenase is secreted by the anaerobic bacterium Clostridium histolyticum, yielding a "crude" collagenase preparation. There are five main types: Collagenase Type I, II, III, IV, and V.

Selection Guide for Collagenase Types

Product Name	Enzyme Activity	Applications
Collagenase I (Type I Collagenase)	≥125 U/mg solid	Epithelial, lung, adipose, adrenal gland tissue dissociation
Collagenase II (Type II Collagenase)	≥125 U/mg solid	Liver, bone, thyroid, heart, salivary gland tissue dissociation
Collagenase III (Type III Collagenase)	≥100 U/mg solid	Mammalian tissue dissociation
Collagenase IV (Type IV Collagenase)	≥125 CDU/mg solid*	Isolation of pancreatic islets, preservation of receptor integrity
Collagenase V (Type V Collagenase)	≥125 CDU/mg solid*	Pancreatic islet isolation, separation of connective tissue into single cells

*CDU = collagen digestion units

Ordering Information

Cat. No.	Product Name	Specification
40507ES60	Collagenase I	100 mg
40508ES60	Collagenase II	100 mg
40509ES60	Collagenase III	100 mg
40510ES60	Collagenase IV	100 mg
40511ES60	Collagenase V	100 mg

2. Organoid Culture Series

2.1 Organoid Growth Media

Introduction

Cebrary™ series organoid growth media are serum-free formulations designed for establishing and maintaining extracellular matrix (ECM)-embedded organoids derived from mouse small intestinal stem cells. The defined composition, including specific cytokines, enables rapid and stable formation of organoids within matrix gel. This product contains no serum.

Product Features

Defined Composition: Chemically defined, serum-free, contains phenol red.

High Safety: Free of toxic or hazardous components; suitable for organoid growth.

Batch-to-Batch Consistency: Strict manufacturing and quality control ensures performance reproducibility.

Low Endotoxin: Sterile-filtered, endotoxin level <2 EU/mL.

Contamination Testing: Confirmed free of mycoplasma, bacteria, and fungi.

High Bioactivity: Potent induction factors promote efficient intestinal organoid differentiation.

Application	Product Name	Cat. No.	Specification
Mouse Small Intestine	Cebrary™ Intestinal Organoid Growth Medium (Mouse)	41426ES60/76	100 / 500 mL
Human Colon Cancer	Cebrary™ Intestinal Cancer Organoid Growth Medium (Human)	41427ES50/60/76	50 / 100 / 500 mL
Human Gastric Cancer	Cebrary™ Gastric Cancer Organoid Growth Medium (Human)	41428ES50/60/76	50 / 100 / 500 mL
Human Liver Cancer	Cebrary™ Liver Cancer Organoid Growth Medium (Human)	41429ES50/60/76	50 / 100 / 500 mL
Human Breast Cancer	Cebrary™ Breast Cancer Organoid Growth Medium (Human)	41430ES50/60/76	50 / 100 / 500 mL
Human Lung Cancer	Cebrary™ Lung Cancer Organoid Growth Medium (Human)	41431ES50/60/76	50 / 100 / 500 mL
Human Esophageal Cancer	Cebrary™ Esophageal Cancer Organoid Growth Medium (Human)	41432ES50/60/76	50 / 100 / 500 mL
Human Ovarian Cancer	Cebrary™ Ovarian Cancer Organoid Growth Medium (Human)	41433ES50/60/76	50 / 100 / 500 mL
Human Pancreatic Cancer	Cebrary™ Pancreatic Cancer Organoid Growth Medium (Human)	41434ES50/60/76	50 / 100 / 500 mL

2.2 Matrix Gel

Introduction

Matrix gel is a soluble basement membrane preparation extracted from Engelbreth-Holm-Swarm (EHS) mouse tumors, rich in extracellular matrix (ECM) proteins. Its major components include laminin, type IV collagen, heparan sulfate proteoglycans (HSPG), and entactin/nidogen. It also contains various growth factors such as TGF-β, EGF, IGF, and FGF. At room temperature, matrix gel polymerizes into a biologically active 3D matrix that mimics the structural, compositional, physical, and functional properties of the in vivo basement membrane, thereby supporting cell culture and differentiation in vitro.

YeaSen Biotechnology has developed Ceturegel™ Matrix LDEV-Free, which is free of lactate dehydrogenase-elevating virus (LDEV), features ultra-low endotoxin levels, and undergoes rigorous mycoplasma testing to ensure absence of contamination. Available in standard concentration, high concentration, and growth factor-reduced variants.

Product Features

High Safety: LDEV-free

Multiple Concentrations: Ranging from 8–20 mg/mL

Batch Consistency: Stringent QC ensures consistent performance

Low Endotoxin: <8 EU/mL

Contamination Testing: Verified free of mycoplasma, bacteria, and fungi

High Batch Yield: Single batch production exceeds 50 L scale

Compatibility: Compatible with all types of cell culture media

Related Products

Product Type	Product Name	Cat. No.	Specification
Standard Concentration	Ceturegel™ Matrix LDEV-Free	40183ES08/10	5 / 10 mL
	Ceturegel™ Matrix Phenol Red-Free, LDEV-Free	40184ES08/10	5 / 10 mL
Low Growth Factor	Ceturegel™ Matrix GFR, LDEV-Free	40185ES08/10	5 / 10 mL
	Ceturegel™ Matrix GFR, Phenol Red-Free, LDEV-Free	40186ES08/10	5 / 10 mL
High Concentration	Ceturegel™ Matrix High Concentration, LDEV-Free	40187ES08/10	5 / 10 mL
	Ceturegel™ Matrix High Concentration, Phenol Red-Free, LDEV-Free	40188ES08/10	5 / 10 mL
	Ceturegel™ Matrix High Concentration, GFR, LDEV-Free	40189ES08/10	5 / 10 mL
For Stem Cell Applications	Ceturegel™ Matrix hESC-Qualified, LDEV-Free	40190ES08/10	5 / 10 mL

2.3 Cytokines

Introduction

The signaling pathways mediating organoid formation mirror those involved in embryonic development and homeostasis in vivo. During culture, various cytokines must be supplemented to activate or inhibit key signaling pathways essential for organoid formation. Different organoid types require distinct combinations of cytokines. YeaSen offers a comprehensive portfolio of organoid-related cytokines covering requirements for organoid initiation, expansion, and differentiation. High bioactivity, low endotoxin levels, and batch-to-batch consistency are critical for successful organoid experiments.

Cytokine Requirements for Different Organoid Types

Organoid Type	Required Cytokines
Stomach	EGF, Noggin, R-spondin 1, Wnt-3a, FGF-4, FGF-10
Intestine	EGF, Noggin, R-spondin 1, Wnt-3a, FGF-2
Liver	EGF, Noggin, R-spondin 1, HGF, FGF-10, FGF-19, BMP-7
Lung	Noggin, R-spondin 1, Activin A, FGF-2, FGF-4, FGF-7, FGF-10
Prostate	EGF, Noggin, R-spondin 1, Activin A, FGF-2, FGF-10, NT-3
Pancreas	EGF, Noggin, R-spondin 1, Wnt-3a, FGF-10
Brain	EGF, Noggin, DKK1, BDNF, GDNF, FGF-2
Mammary Gland	EGF, Noggin, R-spondin 1, Wnt-3a, FGF-2, FGF-10, R-spondin 2, Heregulin β-1
Kidney	BMP-2, BMP-4, BMP-7, FGF-2, FGF-9
Heart	Wnt-3a, Activin A, BMP-4
Retina	Wnt-3a, SHH

Product Features

High Bioactivity: Each batch undergoes functional biological assays.

High Purity: >95% purity (verified by SDS-PAGE and HPLC).

Tag-Free: Native protein sequence without artificial tags.

Low Endotoxin: Most products have <0.01 EU/μg.

Carrier-Free: No BSA or carrier proteins; contains only target protein.

Format: Lyophilized powder for long-term storage stability.

Product Data

High Bioactivity: Protein bioactivity is a key indicator of product quality.

Organoid Case Studies and Results

 Related Products

Protein Name	Species	Cat. No.	Tag	Expression System	Specification
Wnt3a	Human	92276ES60/76	Fc	HEK293	100 / 500 μg
R-Spondin 1	Human	92278ES60/76	None	CHO	100 / 500 μg
R-Spondin 1	Mouse	92277ES60/76	His	CHO	100 / 500 μg
Noggin	Human	92528ES60/76	None	E.coil	100 / 500 μg
Noggin	Mouse	92262ES60/76	None	E.coil	100 / 500 μg
EGF	Human	92701ES60/76	None	Yeast	100 / 500 μg
EGF	Mouse	92703ES60/76	None	E.coil	100 / 500 μg
FGF-2	Human	91330ES60/76	None	E.coil	100 / 500 μg
FGF-2	Mouse	91315ES60/76	None	E.coil	100 / 500 μg
FGF-4	Human	91303ES60/76	None	E.coil	100 / 500 μg
FGF-7	Human	91304ES60/76	None	E.coil	100 / 500 μg
FGF-7	Mouse	91316ES60/76	None	E.coil	100 / 500 μg
FGF-9	Human	91305ES60/76	None	E.coil	100 / 500 μg
FGF-9	Mouse	91318ES60/76	None	E.coil	100 / 500 μg
FGF-10	Human	91306ES60/76	None	E.coil	100 / 500 μg
FGF-10	Mouse	91319ES60/76	None	E.coil	100 / 500 μg
FGF-19	Human	91311ES60/76	None	E.coil	100 / 500 μg
Activin A	Human/Mouse/Rat	91702ES60/76	None	E.coil	100 / 500 μg
BMP-2	Human/Mouse/Rat	92051ES60/76	None	E.coil	100 / 500 μg
BMP-4	Human	92053ES60/76	None	E.coil	100 / 500 μg
BMP-4	Mouse	92056ES60/76	His	E.coil	100 / 500 μg
BMP-7	Human	92054ES60/76	None	E.coil	100 / 500 μg
DKK-1	Human	92275ES60/76	His	HEK293	100 / 500 μg
VEGF165	Human	91502ES60/76	None	Yeast	100 / 500 μg
HGF	Human	92055ES60/76	None	CHO	100 / 500 μg
GDNF	Human	92102ES60/76	None	E.coil	100 / 500 μg
GDNF	Mouse	92103ES60/76	None	E.coil	100 / 500 μg
IGF-1	Human	92201ES60/76	None	E.coil	100 / 500 μg
Shh	Human	92566ES60/76	None	E.coil	100 / 500 μg
Shh	Mouse	92589ES60/76	None	E.coil	100 / 500 μg
BDNF	Human	92129ES60/76	His	E.coil	100 / 500 μg
NRG1	Human	92711ES60/76	None	E.coil	100 / 500 μg
SHH	Human	92566ES60/76	None	E.coil	100 / 500 μg

2.4 Small Molecule Inhibitors and Activators

Introduction

Small molecules are compounds with molecular weights less than 1000 Da (especially <500 Da) that exhibit biological activity. Unlike cytokines and proteins, small molecules can cross cell membranes and exert intracellular effects. They target diverse signaling pathways, regulating cellular processes such as proliferation, differentiation, and self-renewal. These compounds are widely applied in stem cell biology, organoid research, immunology, neurobiology, epigenetics, apoptosis, ion channel studies, oncology, and signal transduction.

Small Molecules Used in Organoid Cultures

Organoid Type	Small Molecule Inhibitors/Activators
Small Intestine	A83-01, Gastrin, Nicotinamide, SB202190, Y27632
Stomach	A83-01, Gastrin, Nicotinamide, SB202190, Y27632
Liver	A83-01, DAPT, Forskolin, Gastrin I, Nicotinamide, Y27632, Prostaglandin E2
Kidney	CHIR99021, Retinoic Acid
Lung	CHIR99021, SB431542
Pancreas	A83-01, Gastrin I, Nicotinamide
Prostate	A83-01, Nicotinamide, SB202190, Y27632, Prostaglandin E2, Testosterone
Mammary Gland	Y27632
Retina	CHIR99021, Y27632
Inner Ear	A83-01, SB431542
Brain	Y27632, MK-2206, GDC-0068, Dorsomorphin

Product Features

High purity and batch-to-batch consistency

Excellent permeability and rapid action

More stable and cost-effective compared to growth factors

Long shelf life and high stability

Effects can be finely tuned via concentration gradients

Rigorously quality-controlled with reliable after-sales support

Quality Control Chromatograms

High purity: majority of products >98%, some >99%

Commonly Used Small Molecule Inhibitors/Activators in Stem Cell/Organoid Culture

Product Name	Cat. No.	Specification	Product Name	Cat. No.	Specification
CHIR-99021	53003ES10	10 mg	Wortmannin	52405ES10	10 mg
SB-431542	53004ES10	10 mg	Trichostatin A	51406ES10	10 mg
LDN193189 (DM-3189)	53012ES10	10 mg	5-Azacytidine	53160ES10	10 mg
LDN-193189 HCl	52606ES10	10 mg	A 83-01	53002ES10	10 mg
DAPT (GSI-IX, LY-374973)	52104ES10	10 mg	Nicotinamide	51402ES10	10 mg
Y-27632	53006ES10	10 mg	AICAR (Acadesine)	52401ES10	10 mg
Y-27632 dihydrochloride	52604ES10	10 mg	Cyclopamine	52913ES10	10 mg
LY-411575 (LY411575)	53014ES10	10 mg	LY294002	52403ES10	10 mg
Valproic acid	52982ES10	10 mg	Prostaglandin (PG) E2	60810ES10	10 mg
Repsox	53016ES10	10 mg	Sodium Butyrate	51501ES10	10 mg
Purmorphamine	53023ES10	10 mg	IDE1	53234ES10	10 mg
Retinoic acid	53001ES10	10 mg	IDE2	53235ES10	10 mg
Forskolin	51001ES10	10 mg	Dorsomorphin dihydrochloride	53010ES10	10 mg
IWR-1	52915ES10	10 mg	SB-202190	53005ES10	10 mg
SB 203580	52002ES10	10 mg	—	—	—

3. Organoid Detection

Introduction

B-27 Serum-Free Supplement (Vitamin A-deficient, 50×) and N-2 Serum-Free Supplement (100×) are the most cited supplements for neural stem cell culture. B-27 is an optimized serum-free supplement that supports the growth and functional maintenance of embryonic, postnatal, and adult hippocampal neurons and other central nervous system (CNS) neurons. N-2 is primarily recommended for neuroblastoma cells and post-mitotic neurons derived from both peripheral (PNS) and central (CNS) nervous systems.

Product Features

Applicability: Suitable for neural stem and progenitor cell culture

Stability: High-quality consistency with strict QC procedures

Sterility Control: Sterile filtered

High Viability Support: Promotes robust cell survival and function

Manufacturing Standards: Produced under controlled conditions ensuring reliability

Related Product Information

Category	Additive	Cat. No.	Specification
Culture Supplements	L-Alanyl-L-glutamine, 200 mM	60701ES60/76	100 / 500 mL
	B-27 Serum-Free Supplement, Vitamin A-deficient, 50×	60704ES10	10 mL
	N-2 Supplement, Serum-Free	60706ES08	5 mL
	Human Transferrin	40133ES60/76	100 / 500 mg
	Insulin, human recombinant	40112ES60/76	100 / 500 mg
	Fetal Bovine Serum Gold (Premium Grade)	40130ES76	500 mL
	L-Glutamine	60314ES60	100 g
	Penicillin-Streptomycin (100×), for cell culture	60162ES76	100 mL
Basal Medium	DMEM/F-12 Reduced-Serum Medium	41420ES76	500 mL
Mycoplasma Series	Mycoplasma Prevention Reagent (2000×)	40608ES03	1 mL
	Mycoplasma Removal Reagent (1000×)	40607ES03	1 mL
	GMyc-PCR Mycoplasma Test Kit	40601ES10/20	10 / 20 assays
	MycAway™ Plus-Color One-Step Mycoplasma Detection Kit	40612ES25/60	25 / 60 T

Organoid Viability Assay

ATP Luminescent Cell Viability Assay Kit

Viability assays measure the physiological and physical health of organoids in response to external stimuli, chemical treatments, or therapeutic interventions, or help determine optimal culture conditions. ATP is the primary energy molecule in cells and reflects metabolic activity, showing a strong linear correlation with live cell number. Therefore, ATP content can be used to quantify viable cells. This kit utilizes a luciferase-based luminescent reaction dependent on ATP to detect intracellular ATP levels, enabling sensitive and wide dynamic range detection of cell viability or live cell counts. It is compatible with low-sample testing as well as high-throughput screening applications.

Figure 12: Schematic diagram of ATP Luminescent Detection Principle

Product Features

1. Simple Operation and Fast Detection

Mix the reagent with the sample at equal volume, incubate for 10 min, then proceed directly to instrument detection—no washing or medium exchange steps required.

2. High Sensitivity and Wide Linear Range

Exhibits excellent linearity across a range from 10 to 100,000 cells. The figures below show comparative testing between Yeasen’s product and a similar product from international competitor P Company.

a. Linearity Test

Figure 13: Demonstrates superior linearity across varying cell numbers and different cell types.

b. Signal Intensity Test

Figure 14: Fluorescence signal is consistently stronger than that of the imported brand (P Company) across different detection time points, with better signal stability.

3. Excellent Stability

Repeated freeze-thaw cycles (up to 5 times) have minimal impact on performance. Stable for 3 days at 4°C with no significant loss. Retains over 80% activity after 1 day at room temperature. Even at ~50% residual activity, the product remains suitable for routine detection applications.

Related Product Information

Type	Product Name	Cat NO.	Size
Viability Assay	ATP Luminescent Cell Viability Assay Kit	40210ES10/60	10/100 ml
Apoptosis Assay	Annexin V-Alexa Fluor 488/PI Apoptosis Detection Kit	40305ES20/50	20/50T
	Annexin V-FITC/PI Apoptosis Detection Kit	40302ES20/50	20/50T
	TUNEL Apoptosis Detection Kit (FITC)	40306ES20/50	20/50T
Proliferation Assay	MTT Cell Proliferation and Cytotoxicity Assay Kit	40206ES76/80	500/1000T

Organoid Immunofluorescence Analysis

Immunofluorescence staining using organoid-specific markers enables assessment of organoid formation, morphology, and structure. Based on the principle of antigen-antibody reactions, immunofluorescence uses fluorescently labeled antibodies and a fluorescence microscope to visualize the location and nature of antigens within tissue or cells. Yeasen offers a wide range of fluorescently labeled secondary antibodies against various species. For low-abundance target proteins that may not be easily detected with standard secondary antibodies, we provide streptavidin-based signal amplification systems. Additionally, to address the issue of fluorescence quenching, we provide mounting media that significantly slow down photobleaching of common fluorophores.

Related Product Information

Application	Product Name	Cat No.	Specification
Antigen Retrieval	Improved Citrate Antigen Retrieval Solution (50X)	36319ES60	100 ml
Detection	Antibody Dilution Buffer	11371ES10/96	10/500 ml
Blocking	Bovine Serum Albumin (BSA), Standard Grade	36101ES25/50	1/10 ml
Control	Normal Goat Serum	36119ES03/10	1/10 ml
Staining	DAPI Stain Solution (5 mg/ml)	40728ES03	1 mg (200 μL)
	Hoechst 33342 Stain Solution (1 mg/mL)	40732ES03/10	1/10 ml
Secondary Antibodies	Alexa Fluor 488-labeled Donkey Anti-Rabbit IgG (H+L)	34206ES60	100 µl
	Alexa Fluor 594-labeled Donkey Anti-Rabbit IgG (H+L)	34212ES60	100 µl
	Alexa Fluor 594-labeled Goat Anti-Rabbit IgG (H+L)	33112ES60	100 µl
	Alexa Fluor™ 488-labeled Goat Anti-Mouse IgG (H+L)	33206ES60	100 µl
	Alexa Fluor 594-labeled Goat Anti-Mouse IgG (H+L)	33212ES60	100 µl
Streptavidin	DyLight 405™-labeled Streptavidin	35102ES60	100 µl
	Alexa Fluor™ 488-labeled Streptavidin	35103ES60	100 µl
	Alexa Fluor™ 647-labeled Streptavidin	35104ES60	100 µl
	Alexa Fluor™ 594-labeled Streptavidin	35107ES60	100 µl
Mounting Media	Fluoromount-G™ Anti-fade Mounting Medium (Water-soluble)	36307ES08/25	5/25 mL
	DAPI Fluoromount-G™ Anti-fade Mounting Medium (Contains DAPI, Water-soluble)	36308ES11/20	4/20 mL

4. Organoid Sequencing and Identification

Introduction

Single-cell sequencing technology utilizes high-throughput next-generation sequencing (NGS) to analyze the sequence information of individual cells, enabling high-resolution insights into cellular heterogeneity and functional roles within the microenvironment.

 Product Features

Related Products

Product Name	Cat No.	Specification
Hieff NGS™ Single Cell/Low Input cDNA Synthesis & Amplification Module	12500ES24/96	24/96T
Hieff NGS™ Single Cell/Low Input RNA Library Prep Kit	12502ES24/96	24/96T
Hieff NGS™ Fast Tagment DNA Library Prep Kit for Illumina™ (for 1 ng)	12206ES24/96	24/96T
Hieff NGS™ DNA Selection Beads	12601ES03/08	1/5 ml

5. Organoid Passaging and Cryopreservation

Introduction

Cebrary™ Cell Recovery Solution for Organoid is designed for the dissociation and passaging of organoids cultured in extracellular matrix (ECM). Its unique formulation enables efficient release of organoids from matrix gel into small cell clusters or single-cell suspensions. The process is gentle and rapid, preserving high cell viability. The product is enzyme-free.

Product Features

Well-defined composition: Free of animal-derived components and enzymes

High safety: Mild reaction, minimal impact on cells and organoids

Excellent batch-to-batch consistency: Strict manufacturing and QC ensure stable performance

Low endotoxin: Endotoxin level < 2 EU/ml

Contamination tested: Confirmed free of mycoplasma, bacteria, and fungi

High production capacity: Batch size over 100 L

Broad compatibility: Compatible with various cell types and organoid systems

Related Products

Product Name	Cat No.	Specification
Cebrary™ Cell Recovery Solution for Organoid	41421ES30/60	30/100 mL
Cebrary™ Cell Freezing Medium for Organoid	41422ES30/60	30/100 mL

6. Compatible Instruments and Consumables

Introduction

 Product Features

High-quality materials: Manufactured from high-transparency imported polystyrene/polypropylene raw materials.

Optimized for cell culture: Cell culture products are TC (tissue culture)-treated to enhance cell adhesion.

Precision pipette tips: Compatible with Gilson, Eppendorf, Sartorius, and other major brands. Tips are straight with smooth inner walls for accurate liquid handling. Available with or without filter plugs.

Cryogenic vials: Temperature resistant from -196°C to 121°C. Sealed cap with silicone gasket prevents leakage. Maximum fill volume should not exceed 80% of the marked capacity. Store in the vapor phase of liquid nitrogen; avoid liquid phase due to risk of vial rupture.

Centrifuge tubes: Smooth inner and outer surfaces with excellent sealing. Clear graduations and white writable area. Temperature resistant from -80°C to 121°C; RCF up to 12,000×g.

Sterile and nuclease-free: Sterile, non-pyrogenic, endotoxin-free, DNase-free, RNase-free.

Product List

Application	Product Name	Cat No.	Specification
Cell Culture Dishes	35 mm Cell Culture Dish	84001ES25	20 pcs/bag, 25 bags/carton
	60 mm Cell Culture Dish	84002ES25	20 pcs/bag, 25 bags/carton
	100 mm Cell Culture Dish	84003ES30	10 pcs/bag, 30 bags/carton
Cell Culture Plates	6-well Cell Culture Plate	84011ES50	1 pc/bag, 50 bags/carton
	24-well Cell Culture Plate	84013ES50	1 pc/bag, 50 bags/carton
	96-well Cell Culture Plate	84015ES51	1 pc/bag, 65 bags/carton
Cell Culture Flasks	Cell Culture Flask, 25 cm², Ventilated Cap	84021ES25	12 pcs/bag, 25 bags/carton
	Cell Culture Flask, 75 cm², Ventilated Cap	84022ES17	5 pcs/bag, 18 bags/carton
	Cell Culture Flask, 175 cm², Ventilated Cap	84023ES10	5 pcs/bag, 10 bags/carton
Cryogenic Vials	Cryogenic Vial, 1.5 mL, Stand-up Bottom, Blue Flat Cap, Sterile	84102ES60	50 vials/pack, 100 packs/carton
Sterile Centrifuge Tubes	CT 15 mL Centrifuge Tube, Sterile, Packed in Bag	83505ES20	25 tubes/bag, 20 bags/carton
	CT 50 mL Centrifuge Tube, Sterile, Packed in Bag	83507ES20	25 tubes/bag, 20 bags/carton
Sterile, Nuclease-free Pipette Tips	10 μL Boxed Sterile Tip	83020ES50	50 boxes/carton
	10 μL Long Filter Tip, Boxed, Sterile	83031ES50	50 boxes/carton
	200 μL Yellow Boxed Sterile Tip	83050ES50	50 boxes/carton
	200 μL Long Filter Tip, Boxed, Sterile	83061ES50	50 boxes/carton
	1000 μL Blue Boxed Sterile Tip	83080ES50	50 boxes/carton
Cell Inserts	24-well Cell Insert, PET/PC Membrane (Transparent), 8 μm, Sterile, TC-treated
Cryopreservation Accessories	Cryogenic Box, Cryogenic Bag

References

[1] Kleinman H K, Martin G R. Ceturegel: basement membrane matrix with biological activity. [C]// Seminars in Cancer Biology. 2005.

[2] http://www.chinazx.org.cn/i.php?id=3398

[3] Corrò, Claudia; Novellasdemunt, Laura; Li, Vivian S.W. (2020). A brief history of organoids. American Journal of Physiology-Cell Physiology, ajpcell.00120.2020.

Appendix

Ceturegel™ Matrix Gel Handling Protocol

How to thaw Ceturegel™ Matrix Gel, and how long does it take?

Place the entire Ceturegel™ vial into a container filled with crushed ice, then place the ice container in a 4°C refrigerator overnight for thawing. Ensure sufficient ice is used. After thawing, gently rotate the vial to check for complete dissolution and absence of ice crystals or gel clumps. We recommend overnight thawing, as high-concentration Ceturegel™ may require extended time. Do not place the Ceturegel™ vial directly into the 4°C refrigerator without ice, and avoid placing the ice container on the refrigerator door or near the door area. Refer to the image below for visual guidance.

What should the appearance of thawed Ceturegel™ Matrix Gel be?

Standard Ceturegel™ Matrix Gel should appear as a clear liquid after thawing. Gel without phenol red is clear and transparent. Higher protein concentrations result in more viscous solutions. Refer to the image below: the two bottles on the left are high-concentration type, and the two on the right are standard type.

How to aliquot and store Ceturegel™ Matrix Gel?

After thawing, Ceturegel™ can be aliquoted into multiple pre-chilled tubes. All aliquoting must be performed using pre-cooled cryotubes, followed by rapid freezing and storage to avoid repeated freeze-thaw cycles. All items in contact with Ceturegel™ (pipettes, tips, tubes) must be pre-chilled before use.

 Why does Ceturegel™ Matrix Gel partially gel during thawing?

Ceturegel™ is highly temperature-sensitive and begins to gel at temperatures above 10°C, with faster gelation above 22°C. If the product arrives in good condition but gels during thawing, this is likely due to improper thawing procedure or temperature fluctuations during the process. Strictly follow the recommended thawing protocol and maintain a consistently low temperature throughout.

How to dilute Ceturegel™ Matrix Gel?

Dilute with pre-chilled serum-free medium or pH 7.4 PBS. Due to batch-to-batch concentration variability, except for product #354277, we recommend diluting to a specific working concentration rather than using a fixed dilution ratio.

What precautions should be taken when using Ceturegel™ Matrix Gel?

All reagents and consumables in contact with Ceturegel™ must be pre-chilled. Handle entirely on ice to maintain low temperature. Ceturegel™ begins to gel once the temperature exceeds 10°C. Refer to the image below for operational guidance.

Cytokine Handling Protocol

1. Centrifugation: The vial contains lyophilized powder. Before opening, centrifuge at 10,000–12,000 rpm for 30 seconds to pellet any protein that may have adhered to the cap or walls.

2. Reconstitution: Dissolve the lyophilized powder in the solvent specified in the product manual. Final concentration should be between 0.1–1.0 mg/mL. Avoid vigorous vortexing, as this may denature the protein. Gently pipette up and down several times to aid dissolution, or allow the solution to sit at room temperature for a period.

3. Dilution: Dilute using PBS or basic culture medium containing 0.1% BSA, 10% FBS, or 5% trehalose (ensure sterility). Use any of the above dilution buffers to further dilute the reconstituted cytokine solution to the desired working concentration. Final concentration should not be lower than 10 µg/mL. Aliquot and store at -20°C to -80°C.

4. Usage: Thaw one aliquot when needed and add directly to your culture medium to achieve the desired working concentration. Avoid repeated freeze-thaw cycles.

Published Customer Articles

Only articles with IF > 5 are listed. Product usage types (cytokines, small molecules, biochemical reagents, cell culture, etc.) are indicated. Only products mentioned in this manual are included. No more than 5 publications per product line.

Cell Culture and Assays

Matrix Gel

[1] Wang Z, Wang Q, Chen C, Zhao X, Wang H, Xu L, Fu Y, Huang G, Li M, Xu J, Zhang Q, Wang B, Xu G, Wang L, Zou X, Wang S. NNMT enriches for AQP5+ cancer stem cells to drive malignant progression in early gastric cardia adenocarcinoma. Gut. 2023 Mar 28:gutjnl-2022-328408. doi: 10.1136/gutjnl-2022-328408. Epub ahead of print. PMID: 36977555. IF: 31.793

[2] Luo K, Gao Y, Yin S, Yao Y, Yu H, Wang G, Li J. Co-delivery of paclitaxel and STAT3 siRNA by a multifunctional nanocomplex for targeted treatment of metastatic breast cancer. Acta Biomater. 2021 Oct 15;134:649-663. doi: 10.1016/j.actbio.2021.07.029. Epub 2021 Jul 18. PMID: 34289420. IF: 19.92

[3] Zhao Y, Lu T, Song Y, Wen Y, Deng Z, Fan J, Zhao M, Zhao R, Luo Y, Xie J, Hu B, Sun H, Wang Y, He S, Gong Y, Cheng J, Liu X, Yu L, Li J, Li C, Shi Y, Huang Q. Cancer Cells Enter an Adaptive Persistence to Survive Radiotherapy and Repopulate Tumor. Adv Sci (Weinh). 2023 Mar;10(8):e2204177. doi: 10.1002/advs.202204177. Epub 2023 Jan 19. PMID: 36658726; PMCID: PMC10015890. IF: 9.0

Cytokines