人骨髓间充质干细胞和汗腺细胞的体外分离与培养*

背景：体外研究人骨髓间充质干细胞和汗腺细胞的分离培养与鉴定,可为探讨骨髓间充质干细胞再生汗腺的可行性打下基础。目的：探寻在体外分离培养骨髓间充质干细胞和汗腺细胞的有效方法。方法：采用直接贴壁法从成人骨髓中体外分离培养骨髓间充质干细胞,并进行扩增和鉴定。采用胶原酶消化法从人全层无烧伤皮肤中分离汗腺细胞,并进行扩增和鉴定。结果与结论：倒置显微镜下见分离培养的骨髓间充质干细胞呈梭形,折光性强,免疫细胞化学染色显示细胞表达CD29、CD105,高表达CD44,不表达造血干细胞表面标志CD34和CD45。汗腺细胞呈扁平多角形,表达汗腺细胞表面标志细胞角蛋白7,8,18,19和癌胚抗原。说明直接贴壁法分离培养骨髓间充质干细胞和胶原酶消化法分离培养汗腺细胞是可行的。

Isolation and culture of human bone marrow mesenchymal stem cells and sweat gland cells in vitro

BACKGROUND: In vitro research on the culture and identification of bone marrow mesenchymal stem cells (BMSCs) and sweat gland cells can provide the foundation for the feasibility of BMSCs regenerating sweat gland. OBJECTIVE: To study the effective method on isolation and culture of sweat gland cells and human BMSCs in vitro. METHODS: The BMSCs were isolated and cultured from adult bone marrow by adherent method, and then expanded and identified in vitro. Sweat gland cells were separated from human full thickness burnless skin using collagenase digestion method, and expanded and identified. RESULTS AND CONCLUSION: The separated and cultured BMSCs showed spindle-shaped and strong refraction under inverted microscope. Immunocytochemistry staining shows that CD29, CD44, CD105 were positive in BMSCs, surface markers CD34 and CD45 of hematopoietic stem cells were negative. Sweat gland cells showed flat polygonal, and surface markers cytokeratin (CK) 7, CK8, CK18, CK19 and carcinoembryonic antigen were positive in sweat gland cells. It is feasible to separate and culture BMSCs with adherent method and to isolate and culture sweat gland cells with collagenase digestion method.