Isolation,Culture and Identification of Endometrial Epithelial Cells of Laying Hens and Establishment of Its Senescence Model
This study aimed to establish a primary culture method for uterine epithelial cells of laying hens and to construct an in vitro senescence model of uterine epithelial cells of laying hens using D-galactose(D-gal).Uterine tissues from egg-laying hens(35 weeks of age)were used as the research object,the uterine epithelial cells of laying hens were isolated and using type Ⅰ collagenase for digestion method,the endometrial epithelial cells from laying hens were isolated and cultured in vitro,the cells were identified by morphological observa-tion and immunofluorescence(IF)staining.The uterine epithelial cells were intervened with different concen-trations of D-gal(0,8,16,32,64 mg/mL),by observed cell morphology and measuring cell viability after 24 hours of intervention for 24 h,and the positive rate of cellular senescence were detected by senescence-asso-ciated β-galactosidase(SA-β-gal)staining,to screen the optimal D-gal intervention concentration.The results showed as follows:1)the cultured uterine epithelial cells of laying hens using scraping combined with enzyme digestion method exhibited a cobblestone-like morphology.2)Immunofluorescence staining using an anti-cy-tokeratin 18(CK18)antibody confirmed the uterine epithelial cells of laying hens,the result was positive,the purity was over 90％.3)Compared with the control group,the cell viability of 8,16,32,64 mg/mL D-gal groups was significantly decreased(P＜0.05),and the cell viability of 32 mg/mL D-gal groups was only 47.87％.4)Compared with the control group,the positive rates of cellular senescence of different concentra-tions of D-gal groups were significantly increased(P＜0.05),and the positive rates of cellular senescence of 16 mg/mL D-gal group was 41.08％.In conclusion,the method used in this study successfully isolated purify and identify the primary chicken uterine epithelial cells,treatment with 16 mg/mL D-gal for 24 h can success-fully construct a senescence model of uterine epithelial cells of laying hens.[Chinese Journal of Animal Nutri-tion,2024,36(12):8041-8048]
endometrial epithelial cellscell culture in vitrosenescence modellaying hens
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维普
