成人乳腺干细胞的培养筛选及生物学特点鉴定

Cultivation and screening of adult mammary gland stem cells and their identifications of biological characters

目的用不同的鉴定方法对应用免疫磁珠技术分离的乳腺干细胞(separated human mammary stem cell,SHMEC)进行生物学特点的鉴定。方法对原代培养的乳腺细胞用免疫磁珠方法(Magnetic Cell Sorting,MACS)进行分离。对分离后的乳腺干细胞(SHMEC)进行免疫组化,光镜,GIEMSA染色,染色体染色,电镜超微结构扫描等方法进行生物学特点的鉴定,并观察其转化为其他乳腺细胞的能力。结果光镜下可见对正常的静止期的乳腺细胞经免疫磁株分离后得到的乳腺导管上皮细胞呈椭圆型,聚集成团。GIEMSA染色可见红色椭圆型细胞及细胞核。扫描电镜下可见细胞的超微结构特点。染色体核型分析染色显示SHMEC细胞为二倍体细胞,染色体数目与正常外周血淋巴细胞相同。染色体恒定,无缺失及短臂丢失。结论从正常组织中经原代培养后分离出的乳腺上皮细胞具有乳腺干细胞的特性并不具有恶性转变的生物学特点。

Objective To identify the biological character of the separated human mammary stem cell （SHMEC） by magnetic cell sorting （MACS） with different methods. Methods The mammary epithelial cells in primary culture were separated by MACS to identify SHMEC＇s biological characters by immunohistochemistry, light microscope, Giemsa staining, chromosome staining and ultramicrostructure scanned by electron microscope and observe the ability of transforming into other mammary cells. Results Under light microscope normal mammary epithelial cells at rest stage separated by MACS turned ellipse and agglomerated. Red ellipse cells and karyons were observed by Giemsa staining. Ultramicrostructures were seen by scanning electron microscope. Chromosome karyotype analysis showed SHMEC were diploid cells, chromosome number were identical with the number of peripheral blood lymphocytes, having chromosome constancy without depletion and gallanconism loss. Condusions The mammary epithelial cells separated from primary culture by MACS have the characters of SHMEC and have no biological features of malignancy transformation.