Cell invasion assays are used to evaluate the ability of cells to migrate through extracellular matrices, a key indicator of metastatic potential. Matrix Gel provides a physiologically relevant basement membrane barrier that cells must degrade and penetrate, thus enabling accurate assessment of invasive behavior under near-physiological conditions.

Figure 1. Migration and Invasion Assay Workflow

I. Protocol for Cell Invasion Assay

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.

II. Experimental Results

Figure 1. Crystal violet staining of cells after invasion assay

III.Troubleshooting

Q1: What is the typical coating density for invasion assays?

A1: A thin coating of 50–100 μL of 1–2 mg/mL Matrix Gel per Transwell insert is commonly used.

Q2: Can the same batch of Matrix Gel be used for both invasion and migration assays?

A2: Yes, the same lot can be used; consistency ensures comparable results across assays.

Q3: How should invasion be quantified?

A3: Staining and microscopic counting or fluorescence-based quantification of invaded cells are both effective.

Q4: What precautions should be taken when handling Matrigel™ (or Ceturegel™)?

A4: All procedures must be performed under sterile conditions. Use pre-chilled pipettes to ensure the matrix remains in a homogeneous, liquid state during handling.

Q5: How should Matrigel™ (or Ceturegel™) be aliquoted and stored?

A5: After thawing, Ceturegel™ matrix can be aliquoted into multiple small tubes. Always use pre-chilled cryovials for aliquoting, freeze rapidly, and store at appropriate temperatures to avoid repeated freeze-thaw cycles. All consumables—including pipette tips, pipettes, and tubes—must be pre-chilled before use to maintain matrix stability.

Related Products

Category	Name	Cat. No.	Size	Note
Basic concentration of Matrigel 8-12 mg/mL	Ceturegel™ Matrix LDEV-Free	40183ES08/10	5 mL / 10 mL	Suitable for 2D/3D culture, invasion and migration assays, and in vivo tumor formation studies.
	Ceturegel™ Matrix Phenol Red-Free，LDEV-Free	40184ES08/10	5 mL / 10 mL
High concentration 18-20 mg/mL	Ceturegel™ Matrix High Concentration，LDEV-Free	40187ES08/10	5 mL / 10 mL	High-concentration gel is viscous and gels quickly, ideal for in vivo tumor formation with hard-to-grow cell lines.
	Ceturegel™ Matrix High Concentration，GFR，LDEV-Free	40189ES08/10	5 mL / 10 mL
	Ceturegel™ Matrix High Concentration，Phenol Red-Free，LDEV-Fre	40188ES08/10	5 mL / 10 mL
Low Growth Factor	Ceturegel™ Matrix GFR, LDEV-Free	40185ES08/10	5 mL / 10 mL	Minimizes growth factor interference in signaling pathway studies.
	Ceturegel™ Matrix GFR, Phenol Red-Free, LDEV-Free	40186ES08/10	5 mL / 10 mL
Stem cell	Ceturegel™ Matrix hESC-Qualified，LDEV-Free	40190ES08/10	5 mL / 10 mL	Mainly used for hESC/iPSC stem cell culture
Organoid-Specific	Ceturegel™ Matrix for Organoid culture, Phenol Red-Free, LDEV-Free	40192ES08/10	5 mL / 10 mL	Upgraded organoid matrix gel for normal and tumor tissues with improved culture performance.