山羊子宫内膜基质细胞的分离培养及鉴定

Isolation, culture and identification of goat endometrium stromal cells

对山羊子宫内膜基质细胞进行分离与体外培养,研究其生长特性,并对其进行免疫细胞化学染色鉴定。结果表明,山羊子宫内膜在37℃条件下,以2 g／L的胶原酶Ⅰ型消化3～4 h效果最好;采用过滤—低速离心—沉降相结合的方法分离山羊子宫内膜基质细胞效果较好,所得细胞在培养后0．5 h开始贴壁,培养12 h后,可见梭形和多角形2种形态的细胞,3～4 d呈网状铺满皿底,有明显的重叠生长现象;免疫细胞化学染色显示这2种形态的细胞均表达波形蛋白,阳性率可达95％以上,不表达角蛋白。说明采用本文所述的消化和分离方法可获得高纯度的子宫内膜基质细胞。

This paper focuses on optimal condition of the isolation and vitro culture of stromal cells of endometrium in goat, and studies the growth characteristics and immunocytochemical staining. The results showed that the tissues presemted best digested with Collagenase Ⅰ of 2 g/L at 37 ℃ 3-4 h ; stromal cells were isolated by filtering-brachytely centrifugalization-sedimentation, and became adherent after 0.5 h of culture, two shapes of cells with the forme of spindle and polygonal, were found after 12 h of culture. Immunocytochemical staining method showed that this 2 shapes of stromal cells were positive for vimentin, with 95% of positive rate, and negative for cytokeratin. The conclusion can be drawn that high purified stromal cell can be obtained by the digestion and separation method in suggested this article.