小鼠乳腺上皮细胞的体外培养

采用I型、II型胶原酶和胰蛋白酶混合（1：1：1）消化乳腺组织，并用差速贴壁法纯化培养乳腺上皮细胞，倒置显微镜下观察细胞的形态，并用角蛋白18免疫荧光染色的方法对细胞进行鉴定。结果表明：改良型混合酶消化法能获得较多的细胞团，且12h后细胞团绝大多数已贴在细胞瓶壁，72h后细胞团完全铺开呈单层融合，细胞呈典型的铺路石样。免疫荧光鉴定结果显示，角蛋白18呈阳性反应。因此，应用改良型混合酶消化法能在短时间内获得大量较纯的乳腺上皮细胞。

In vitro Culture of Mouse Mammary Epithelial Cells

The mixture （ 1： 1：1 ） of collagenase types I, II and trypsin was used to dissociate breast tissue,consequently epithelial ceils were purified and cultured with differential cell adhesion method. The cell growth was evidenced by revealing the morphological characteristics under inverted microscope, and detected by Keratin 18 using indirect immunocytochemistry method. The results showed that epithelial organoids could be mostly acquired and had attached to the plastic dish after 12 h, then ceils spread and formed confluent monolayer of cobblestone, which might last for 60 h. In addition, the tissue - specific expression of cytokeratin 18 in mammary epithelial cells was identified via cytokeratin immunohistochemistry. Therefore, plenty of pure mammary epithelial ceils may be harvested by improved collagenase/trypsin digestion.