METHOD FOR ISOLATING FIBROBLASTS FROM THE CORNEAL STROMA

摘要

FIELD: cell biology and medicine.;SUBSTANCE: invention relates to cell biology and medicine, in particular to a method for isolating fibroblasts from the corneal stroma. To implement the method, stromal lenticules obtained by laser vision correction are used. Crushed lenticules are treated with a 0.05-0.1% solution of type I collagenase, with the addition of 2% fetal bovine serum (FBS) at 37°C for 18-20 hours. Then, the debris is precipitated from the cell suspension by centrifugation, the supernatant is collected, the cells are precipitated from it by centrifugation, the cells are washed twice with a buffer solution. Then the cell pellet is resuspended in DMEM/F12 medium and cultured in DMEM/F12 medium supplemented with 10% FCS, 2 mM L-glutamine, 40 mcg/ml gentamicin. Then the cells are cultured at 37°C and 5% CO2 until the required cell passage is achieved.;EFFECT: invention makes it possible to simplify the method for obtaining cells and improve the quality of the target product.;2 cl, 2 ex