动物造模 z-bio 2021-11-12 416

　　Objective: To explore a simple and easy method to isolate and culture tree shrew corneal stromal cells in vitro quickly.

　　Methods: The experiment was divided into experimental group A and experimental group B. The corneal stroma of the tree shrew was taken, and the experimental group A was cut into pieces, digested with 1% type II collagenase for 60 minutes, centrifuged and resuspended and inoculated; experimental group B used type II collagen Enzyme combined with neutral protease digestion. Compare and record the growth of primary and passage cells. Passage time. Cells are identified by vimentin immunohistochemistry and immunofluorescence.

　　Results: The collagenase digestion method can successfully obtain the primary cells, which can be passaged after 9 days, and the cell morphology is good; the passaged cells grow faster and can be passed down to one generation after 2 to 3 days; the double enzyme digestion method can obtain fewer CSCs, 10 days Only a small amount of scattered cells can be seen, which can be passaged in 15-20 days; vimentin immunohistochemistry and immunofluorescence expression are positive.

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