摘要
[目的]建立牦牛子宫内膜腺上皮细胞体外分离培养方法。[方法]分别采用组织块培养法和消化培养法分离、纯化牦牛子宫内膜腺上皮细胞。[结果]组织块培养约8 d细胞可汇合成片,经胰蛋白酶分次消化,可得到纯化的子宫内膜上皮细胞;使用2 g/L的胶原酶Ⅱ消化2.5 h,更换新鲜消化液继续消化2.5 h,消化悬液经74μm滤网过滤,400 r/min离心5 min,收集沉淀,重悬后自然沉降,可得到纯净的腺细胞团。免疫细胞化学显示所得细胞角蛋白染色阳性,阳性率达到95%以上。[结论]2种方法均可得到纯净的牦牛子宫内膜上皮细胞。
[ Objective] To develop in-vitro culture method of yak endometrial gland epithelial cells. [ MethodJ The gland epithelial cells were i- solated from yak endometrium by explant culture method and digestion culture method, respectively. [ Result ] In the first method, the ceils isolated from the endometrium explant could merge into a single layer after 8-d culture, and they coukt be purified by gradation digestion with trypsase. In the second method,the endometrium explant were first digested by collagenase lI by incubation at 37℃ for 2.5 h and then further digested in fresh 2 g/L collagenase ]I for another 2.5 h. The cell suspension was leached through 74μm filter and centrifuged at 400 r/min for 5 min. Then the cell pellet was re-suspended,followed by natural sedimentation to collect purified gland epithelial ceils. The isolated ceils were cytokeratin- positive as detected by immunocytochemical staining,and the positive rate could reach 95%. [ Conclusion] The yak endometrial gland epithelial cells can be isolated and purified by both the explant cuhure method and digestion culture method.
出处
《安徽农业科学》
CAS
北大核心
2011年第18期11085-11087,11110,共4页
Journal of Anhui Agricultural Sciences
基金
教育部科学技术研究重点项目"(210216)
"211工程"三期高层次人才科研启动费项目(SZRC-211-05)
关键词
牦牛
子宫内膜
腺上皮细胞
体外培养
Yak
Endometrium
Gland epithelial cells
In-vitro culture
