奶山羊乳腺上皮细胞的体外培养及形态观察

Study on the cultivation and morphological observation of dairy goat's mammary gland epithelial cells in vitro

【目的】建立奶山羊乳腺上皮细胞体外培养方法,并对原代和传代细胞进行形态观察。【方法】分别采用组织块法和胶原酶消化法培养奶山羊原代乳腺上皮细胞,然后通过胰酶消化法结合反复贴壁法纯化细胞,免疫组化法鉴定细胞,并利用倒置显微镜观察细胞形态。【结果】组织块法和胶原酶消化法均能培养奶山羊原代乳腺上皮细胞;纯化获得的细胞呈卵圆形或多角形,形成典型的鹅卵石或铺路石样,且细胞角蛋白检测呈阳性;多次传代后乳腺上皮细胞出现分化现象,形态多样,增殖仍然旺盛。【结论】成功建立了一种简单易行、经济而实用的奶山羊乳腺上皮细胞体外培养方法。

【Objective】 The aim of this paper was to establish a method of the dairy goat mammary gland epithelial cells in vitro,and to observe the morphology of the cultured primary and passage cells.【Method】 Two cultural methods of the tissue and collagenase digestion were respectively used for culturing the primary mammary gland epithelial cells in vitro.The method of the trypsinization combining the repeat adherence was utilized for purifying the cells,and the immunohistochemistry method was used for identifying the cells.At the same time the morphology of the primary and the passage cells was observed by the inverted microscope.【Result】 The primary cells also could be cultured by the tissue and collagenase digestion cultural methods.The purified cells were orbicular-ovate or polygon,forming a typical cobblestone-like or slab stone-like appearance,and the cytokeratin detection was positive.After the high passage the cells could be differentiated into many morphous varieties,and the proliferative ability was still high.【Conclusion】 The culture method of the diary goat mammary gland epithelial cells was constructed in vitro.This method was simple,maneuverable,economic and pragmatic.