Organoid culture has become increasingly popular in recent years. Organoids are derived from adult or pluripotent stem cells differentiated in vitro, capable of mimicking tissue/organ architecture and function regarding cell types, spatial organization, and physiology. Advancements in organoid technologies have propelled development in developmental biology, genetics, pathology, toxicology, and more.
Basic Knowledge
Do organoid cultures require specific sources of starting cells?
Yes. Most organoids originate from epithelial cells. Culture strategies for organoids of non-epithelial origin require further development.
Are organoids composed of only a single cell type?
No. Organoids are grown from stemlike starting cells that proliferate and differentiate, self assembling into structures containing multiple cell types.
Can organoids be cryopreserved and revived like cell lines?
Yes. Cryopreservation is typically performed after 2–3 passages, but yields improve when organoids are matured (after 7–10 passages).
Experimental Operations
Can you use cryopreserved tissue instead of fresh tissue to derive primary cells for 3D culture?
Yes, but the tissue chunk size must be appropriate. Cell viability in cryopreserved samples is significantly reduced, lowering the success rate in downstream culture.
What is the standard for passaging organoids?
First passage typically occurs at days 7–14, followed by subsequent passaging every 7–10 days.
Is size control important in organoid culture?
Yes. Organoids should be kept under ~500 µm. Without vascularization or gas exchange, cells near the core may suffer from nutrient and oxygen deprivation, leading to necrosis.
Identification and Application
How are organoids characterized?
Morphological inspection via microscopy and H&E staining
Expression analysis of biomarkers using Western blot, qRT-PCR, immunofluorescence, and flow cytometry
Genomic sequencing to detect feature loss
Functional assays, depending on the organoid type
When are organoids suitable for drug testing?
Use early-passage (P2–P3) fresh organoids to minimize culture artifacts.
For thawed organoids, re-culture at least one passage before testing, as morphology can shift post-thaw.
Can organoids be transplanted in vivo for organ reconstruction?
Animal experiments show feasibility, but clinical applications have not yet materialized.
Other Points
How does organoid culture differ from traditional cell culture?
Source cells are stem-like epithelial cells
Culture requires extracellular matrix (e.g., basement-membrane gels) for 3D structure
In vitro differentiation and self-organization necessitate complex cytokine cocktails
How do you prove the structure is a true organoid and not just a multicellular aggregate?
Organoids mimic in vivo organ development and display organized architecture and spatial biomarker distribution, unlike simple aggregates. Immunofluorescence of marker localization provides clear evidence.
Product Overview
|
Product Name |
Specification |
Cat No. |
|
CHIR-99021 |
2 mg/5 mg/10 mg |
53003ES05/08/10 |
|
Ceturegel™ Matrix High Concentration, LDEV-Free |
5 mL/10 mL |
40187ES08/10 |
|
Ceturegel™ Matrix GFR, LDEV-Free |
5 mL/10 mL |
40185ES08/10 |
|
Recombinant Human IL-2 Protein |
10 μg/50 μg/100 μg/1 mg |
90213ES10/50/60/80 |
|
Recombinant Human IL-10 Protein |
2 μg/10 μg/50 μg/100 μg/500 μg |
90109ES05/10/50/60/76 |