How can fibroblast contamination be reduced in epithelial cell preparations?
Fibroblast contamination cannot be completely avoided in primary epithelial cell cultures. However, it can be effectively reduced by taking advantage of the different adhesion properties of epithelial cells and fibroblasts.
Recommended approach: partial trypsinization
1. Add trypsin/EDTA to the culture dish.
2. Incubate for 2-4 minutes.
3. During this time, fibroblasts detach first.
4. Inactivate the enzyme once fibroblasts are detached.
5. Aspirate the cell suspension containing the fibroblasts.
6. Wash the remaining epithelial cells twice with buffer.
7. Continue culturing the epithelial cells in the appropriate Growth Medium.
Recommended approach: partial trypsinization
- Epithelial cells attach more firmly to tissue‑culture plastic than fibroblasts.
- This difference allows selective removal of fibroblasts by short trypsinization.
1. Add trypsin/EDTA to the culture dish.
2. Incubate for 2-4 minutes.
3. During this time, fibroblasts detach first.
4. Inactivate the enzyme once fibroblasts are detached.
5. Aspirate the cell suspension containing the fibroblasts.
6. Wash the remaining epithelial cells twice with buffer.
7. Continue culturing the epithelial cells in the appropriate Growth Medium.
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