Introduction of culture methods for epithelial cells

Epithelial cells, including glandular epithelium, are functional components of many organs such as liver, pancreas, and mammary glands. Because cancer originates from epithelial tissues, epithelial cell culture is particularly valued. However, fibroblasts are often mixed in epithelial cell culture, and growth rate during culture. It often exceeds epithelial cells and is difficult to purify. At the same time, epithelial cells are difficult to survive in vitro, so purification and prolonged survival are the key to culture.
The epithelial cells in the body grow on the basement membrane composed of collagen, so the culture may be conducive to growth on the substrate with collagen, and the human or mouse epidermal cells are cultured in the 3T3 cells as the feeder layer (after irradiation with radiation), the cells are easy to grow. A certain degree of differentiation can occur. Lowering PH, Ca2+ content and temperature, and adding epidermal growth factor to the medium are beneficial to the growth of epidermal cells.

Taking epidermal cells as an example, pure epithelial cells can be obtained by skin epidermal and dermal isolation culture methods, and the method is as follows (Blackwood Stool 1981)
1. Material: Surgical skin graft or surgical residual skin, the skin of premature abortion is better, and the keratinized layer is thin, cut into 0.5-1 cm cm small pieces.
2, set 0.02% EDTA, room temperature, 5 minutes.
3. Change to 0.25% trypsin and overnight at 4 °C.
4. Separation: remove the skin and separate the epidermis from the dermis with a vascular clamp or forceps.
5, remove the epidermis, cut into smaller pieces, set 0.25% trypsin, 37 ° C, 30-60 minutes.
6, repeated blowing, made into a suspension.
7. Culture: After filtering with 80 mesh stainless steel gauze, centrifuge at low speed and aspirate the supernatant.
8. Directly add the medium (Eagle plus 20% calf serum) to make a cell suspension, inoculate the flask, and incubate in a CO2 incubator.