This protocol is based on the method of Ehaio et al. (1987) and can be modified for the isolation and culture of other epithelial cells if tissue is available. Fresh bovine eyes were obtained from the abattoir and immediately placed on moist ice. This experiment was obtained from the Cellular Experiment Guide (previous volume) by Huang Peitang.

Operation method

Isolation and culture of bovine keratinized epithelial cells

Move

1) Wash the bull's eye with 1% iodine polyvinylpyrrolidone solution, rinse with water, and treat briefly with 70% ethanol.

2) A 3.5 mm biopsy drill was used to excise a small piece of epithelial tissue from the eye together with the connective tissue underneath (preferably the dendritic corneal limbal tissue).

3) The tissue was placed on a dry polylysine-coated surface and allowed to adhere for a full 10 minutes.

4) Add culture medium (MEM, containing 10% FCS, 100IU/ml penicillin, 100ug/ml streptomycin, 2~4 mmol/L L-glutamine) to the tissues and incubate at 37°C for 2 days.



5) Remove the tissue mass carefully with sterile forceps.

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