动物造模 z-bio 2022-03-09 573

　　OBJECTIVE: To explore a simple and easy method for rapid isolation and culture of tree shrew corneal stromal cells in vitro.

　　Methods: The experiment was divided into experimental group A and experimental group B. The corneal stroma of tree shrew was taken, the experimental group A was cut into pieces, digested with 1% type II collagenase for 60 min, centrifuged and resuspended for inoculation; the experimental group B was treated with type II collagen. Enzyme combined with neutral protease digestion. The growth and passage time of primary and passaged cells were compared and recorded, and the cells were identified by vimentin immunohistochemistry and immunofluorescence.

　　Results: The collagenase digestion method can successfully obtain primary cells, which can be passaged after 9 days, and the cell morphology is good; the passaged cells grow faster, and can be passaged in 2-3 days; the double-enzyme digestion method can obtain less CSCs, 10 days Only a few scattered cells can be seen, and they can be passaged at 15-20 days; the expression of vimentin immunohistochemistry and immunofluorescence is positive.

　　Conclusion: Both methods can successfully culture CSCs, but the collagenase digestion method can more easily and efficiently isolate and culture a large number of tree shrew CSCs.