The aim of this experiment was to establish a primary culture method of equine mammary epithelial cells to provide experimental materials and cell models for the study of lactation-related mechanisms in equine animals.The experiment was conducted on mammary tissue of Kazakh mares in lactation,and equine mamma-ry epithelial cells were isolated using type Ⅰ collagenase and hyaluronidase,and purified and characterized.The results showed as follows:1)the morphology of equine mammary epithelial cells cultured by enzyme di-gestion method was"strip-like"or"island-like";mammary epithelial cells cultured in resuscitation after freez-ing were in good growth condition.2)The purified equine mammary epithelial cells had a normal"S"shape growth cycle.3)Agarose gel electrophoresis results showed that the cultured equine mammary epithelial cells were able to express their specific gene cytokeratin 18 gene(CK1 8).4)The mouse anti-cytokeratin 18 was used as primary antibody to identify the equine mammary epithelial cells by immunofluorescence technique,and the result was positive.In conclusion,the method used in this study is able to successfully isolate,purify culture and identify equine mammary epithelial cells.[Chinese Journal of Animal Nutrition,2023,35(12):8046-8052]
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