中文摘要：

目的鉴定人脐带沃顿胶间充质干细胞（hUCWJ-MSCs）经汗腺诱导培养基培养后向汗腺样细胞分化的潜能。方法酶消化法分离培养人正常汗腺细胞,热休克汗腺细胞后收集培养基上清液,按10%的体积分数配制汗腺分化诱导培养基。酶消化法分离获取hUCWJ-MSCs,流式细胞仪分析细胞表型特征;成骨诱导培养基和成脂诱导培养基中培养2～3周后采用碱性磷酸酶染色和油红-O染色对其分化潜能进行鉴定;在汗腺诱导培养基中培养3周,倒置显微镜观察细胞形态变化,分别收集培养1、2、3周时的细胞,采用免疫组织化学和流式细胞术检测汗腺标记物CEA、CK14、CK19的表达,RT-PCR检测汗腺发育基因（EDA）的表达。结果正常汗腺细胞呈铺路石状克隆样增生。hUCWJ-MSCs呈梭形、成纤维细胞样,流式细胞分析显示其CD44、CD105、CD34、CEA阳性率分别为97.37%9、6.26%、0.56%0、.52%;经成骨诱导培养基和成脂诱导培养基培养2～3周后,可诱导成为油红-O染色阳性的脂肪细胞和碱性磷酸酶染色阳性的骨细胞;在汗腺诱导培养基中培养3周后,细胞具有类似汗腺样结构,免疫组织化学DAB显色结果显示分化后的hucwJMSCs表达汗腺细胞标记物CEA、CK14、CK19，流式细胞仪检测其阳性率分别为54．37％、60．25％、62．13％，RTPCR结果显示其可较高水平地表达EDA基因。结论hUCWJ—MSCs具有向汗腺样细胞分化的潜能。

英文摘要：

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 10% volume fraction.hUCWJ-MSCs were harvested by enzyme digestion,and the cell phenotypes were analyzed by flow cytometry（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,CK14 and CK19） 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,CD105,CD34 and CEA detected by 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,CK14 and CK19 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 s