Comparison Of The Effects Of Four Enzyme Mass Concentrations On The Isolation Of Mouse Testicular Interstitial Cells
Keywords
Leydig Cells, Trypsin, Collagenase Ii, Lipid Droplets, Mice
Abstract
Abstract Purpose: To compare the separation effects of four enzyme mass concentrations on primary mouse testicular Leydig cells. Methods: The testicles of 20 adult male Kunming mice were evenly divided into A, B, C and D4 groups, and were digested and separated with 0.05%, 0.125%, 0.25% trypsin and 0.02% collagenase II respectively. Testicular Leydig cells, compare the number and survival rate of Leydig cells separated by four enzyme mass concentrations; observe the Leydig cells and their lipid droplet morphology using hematoxylin-eosin staining and Sudan III staining respectively. Results: The number of isolated interstitial cells in groups A and B was more than that in groups C and D (P<0.05). The survival rates of interstitial cells isolated by the four enzyme mass concentrations were all above 90%, and the difference was not statistically significant (P>0.05). The mesenchymal cells isolated by low concentrations of trypsin (0.05% and 0.125%) were in good growth status, with small and abundant intracellular lipid droplets. After long-term in vitro culture, the cytoplasm became vacuolated and the lipid droplets fused. Conclusion: Low mass concentration of trypsin is effective in isolating primary stromal cells.
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Original research was done by Huang Rui, Zhu Weijie
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