解放军医学杂志2011，Vol.36Issue(3)：211-214,4.
特定微环境诱导人脐带沃顿胶间充质干细胞向汗腺样细胞分化的实验研究
Differentiation of human umbilical cord Wharton's jelly derived mesenchymal stem cells into sweat gland-like cells under special microenvironment: an experimental study
摘要
Abstract
Objective To identify the potentiality of human umbilical cord Wharton's jelly derived mesenchymal stem cells (hUCWJ-MSCs) differentiated into sweat gland-like cells under the cultivation of sweat gland-induction medium.Methods Sweat gland cells were harvested from normal skin by digesting with collagenase typeⅡ. heat-shocked and then the supernatants of medium were collected. The sweat gland-induction medium was prepared at lO％ volume fraction. hUCWJ-MSCs were harvested by enzyme digestion, and the cell phenotypes were analyzed by flow cvtometry (FCM). Alkaline phosphatase (ALP) and Oil red-O staining were then performed after culturing in osteogenic and adipogenic induction medium for 2 - 3 weeks respectively. The hUCWJ-MSCs were cultured in sweat gland-induction medium for 3 weeks, the changes of cell morphology were observed with inverted microscope; the cells cultured for 1 , 2 and 3 weeks were harvested. and the expression of sweat gland markers (CEA, CKl4 and CKl9) were determined by immunohistochemistry and FCM, the expression of sweat gland development gene (EDA) was determined by RT-PCR. Results The normal sweat gland cells exhibited clonal growth with a flagstone appearance. while the hUCWJ-MSCs showed spindle and myofibroblast-like phenotype, and the positive rate of CD44 , CDl05 , CD34 and CEA detected bv FCM was 97.37％, 96.26 ％, . 0.56％, and 0.52％, respectively. After cultured in osteogenic and adipogenic induction medium for 2-3 weeks. the hUCWJ-MSCs were induced into adipocytes of Oil red-O positive staining and osteocytes of ALP positive staining. respectively. After cultured in sweat gland-induction medium for 3 weeks, sweat gland-like structures were found, and sweat gland markers CEA, CKl4 and CKl9 were positive in differentiated hUCWJ- MSCs when detected by immunohistochemistry, the positive rate detected by FCM was 54.37％, 60.25 ％, and 62.13 ％ , respectively. RT-PCR analysis revealed a high level expression of gene EDA in differentiated hUCWJ-MSCs. Conclusion The hUCWJ- MSCs has a potentiality of differentiation into sweat gland-like cells.关键词
汗腺/脐带/间质干细胞/细胞分化/再生医学分类
医药卫生引用本文复制引用
许永安,黄沙,付小兵,马奎,张翠萍..特定微环境诱导人脐带沃顿胶间充质干细胞向汗腺样细胞分化的实验研究[J].解放军医学杂志,2011,36(3):211-214,4.基金项目
国家重点基础研究发展计划(2005CB522603) （2005CB522603）
国家自然科学基金重点项目(30672176,30730090) （30672176,30730090）
全军"十一五"攻关课题(06G116) （06G116）
