Bu Zhang-Yu, Wu Li-Min, Yu Xiao-Hong, Zhong Jian-Bo, Yang Ping, Chen Jian
Department of Dermatology, Hangzhou First People's Hospital, Nanjing Medical University, Hangzhou, Zhejiang 310006, P.R. China.
Exp Ther Med. 2017 Jul;14(1):303-307. doi: 10.3892/etm.2017.4456. Epub 2017 May 16.
The present investigation explored the culture, isolation and characterization of hair follicle cell differentiation from umbilical cord blood mesenchymal stem cells (MSCs). Flow cytometry was used to obtain MSCs from the isolation and purification of human umbilical cord blood MSCs. Culture suspension of hair follicle organ was centrifuged and the supernatant used in the culture medium of MSCs, and the entire process of induced differentiation was recorded by photomicroscopy. The expression level of surface marker CK15 of hair follicle cells obtained from induced differentiation was detected with immunofluorescence. RT-PCR method was used to further detect the difference in expression of CK15 between hair follicle cells and umbilical cord blood MSCs, and statistical analysis was carried out. CD44CD29 double-labeled cells accounted for 50.8% of all the samples of umbilical cord blood MSCs in this study. The diameter of hair follicle cells differentiated from umbilical cord blood stem cells reached 800×10 mm after 3 weeks of cell culture. Based on the detection and colocalization of CK15 expression in induced hair follicle cells, the overlap ratio between CK15 and nuclei reached 83% in hair follicle cells, which was obviously higher than that in umbilical cord blood stem cells. The difference had statistical significance (P<0.05). In conclusion, hair follicle cells can be successfully differentiated from umbilical cord blood stem cells by using the supernatant from hair follicle cells. This method can be used for high-speed induced differentiation with high purity, which is promising for clinical application.
本研究探讨了脐血间充质干细胞(MSCs)向毛囊细胞分化的培养、分离及鉴定。采用流式细胞术从人脐血MSCs的分离纯化中获取MSCs。将毛囊器官培养悬液离心,所得上清液用于MSCs的培养基中,并通过光学显微镜记录诱导分化的全过程。用免疫荧光法检测诱导分化所得毛囊细胞表面标志物CK15的表达水平。采用RT-PCR法进一步检测毛囊细胞与脐血MSCs中CK15表达的差异,并进行统计学分析。本研究中,CD44CD29双标记细胞占脐血MSCs所有样本的50.8%。脐血干细胞分化的毛囊细胞在细胞培养3周后直径达到800×10 mm。基于诱导毛囊细胞中CK15表达的检测及共定位,毛囊细胞中CK15与细胞核的重叠率达到83%,明显高于脐血干细胞,差异具有统计学意义(P<0.05)。综上所述,利用毛囊细胞的上清液可成功将脐血干细胞诱导分化为毛囊细胞。该方法可用于高纯度的快速诱导分化,具有良好的临床应用前景。