In vitro matrigel fibroblast invasion assay

Area of Science:

• Biomedical Sciences
• Cell Biology
• Rheumatology

Background:

• Rheumatoid arthritis (RA) involves joint inflammation and cartilage degradation.
• Fibroblast-like synoviocytes (FLS) play a key role in RA pathogenesis through cartilage invasion.
• Quantifying FLS invasiveness is crucial for understanding RA progression.

Purpose of the Study:

• To develop and validate an in vitro invasion assay for fibroblast-like synoviocytes (FLS).
• To provide a quantitative method for assessing FLS invasiveness in a controlled laboratory setting.
• To facilitate research into the mechanisms of cartilage degradation in rheumatoid arthritis.

Main Methods:

• An in vitro invasion assay using transwell inserts coated with collagen IV and matrigel.
• Seeding of FLS in serum-free medium within the transwell.
• Incubation for 3 days followed by fixation, staining, and microscopic counting of invaded cells.

Main Results:

• The assay effectively models FLS growth through an artificial extracellular matrix.
• Paraffin coating of transwell walls prevents meniscus formation, ensuring assay accuracy.
• Quantifiable data on FLS invasion can be obtained via light microscopy and cell counting.

Conclusions:

• The developed in vitro assay is a reliable tool for measuring FLS invasiveness.
• This assay aids in studying the cellular mechanisms underlying rheumatoid arthritis.
• Further research can utilize this method to explore therapeutic strategies targeting FLS invasion.