人尿源性干细胞的原代培养

doi:10.3969/j.issn.1004-616x.2017.05.014

癌变·畸变·突变 ›› 2017, Vol. 29 ›› Issue (5): 390-393.doi: 10.3969/j.issn.1004-616x.2017.05.014

人尿源性干细胞的原代培养

周明, 孙震晓

北京中医药大学生命科学学院, 北京 100029

收稿日期:2017-05-30 修回日期:2017-08-18 出版日期:2017-09-30 发布日期:2017-09-30
通讯作者: 孙震晓,E-mail:sunzxcn@hotmail.com E-mail:sunzxcn@hotmail.com
作者简介:周明,E-mail:737194066@qq.com。

Cultivation of human urine-derived stem cells

ZHOU Ming, SUN Zhenxiao

Life Science College, Beijing University of Chinese Medicine, Beijing 100029, China

Received:2017-05-30 Revised:2017-08-18 Online:2017-09-30 Published:2017-09-30

摘要/Abstract

摘要： 目的：建立一种人尿源性干细胞（hUSCs）的原代分离培养及保存方法，为hUSCs的应用奠定基础。方法：取10例成人健康志愿者的尿液分别进行hUSCs原代培养，台盼蓝染色法比较不同冻存液配方复苏后的细胞存活率，进而确定hUSCs的最佳冻存液配方；采用四甲基噻唑蓝（MTT）法检测hUSCs细胞的增殖活性；流式细胞术测定间充质干细胞表面抗原CD44和CD90的表达情况。结果：10例健康成人志愿者中6例成功提取了hUSCs并体外培养形成集落；台盼蓝染色法确定配比为DMSO∶FBS∶hUSCs培养基=1∶4∶5的冻存液细胞存活率最高，与其他组比较差异具有统计学意义（P < 0.05）；MTT法检测显示hUSCs生长曲线呈S形且具有良好的增殖活性；流式细胞术鉴定干细胞表面抗原CD44和CD90呈阳性表达。结论：成功建立了一种人尿源性干细胞的原代分离培养及保存方法。

关键词: 人尿源性干细胞, 原代培养, CD44, CD90

Abstract: OBJECTIVE: To establish a method for isolation,cultivation and preservation of human urine-derived stem cells(hUSCs). METHODS: hUSCs were isolated from fresh urine samples from adult healthy volunteers. Survival rate of hUSCs after cryopreservation and resuscitation was detected by trypan blue staining to determine the best formula for freezing medium. Proliferation of hUSCs was determined by MTT assay. Stem cell surface marks were detected by flow cytometry. RESULTS: 6/10 hUSCs were successfully isolated and cultured from urine. The survival rate of hUSCs in the freezing medium-DMSO:FBS:hUSCs medium=1:4:5-was the highest based on trypan blue staining. MTT assay showed that the growth curve of hUSCs was S-shaped showing good proliferation activity. Mesenchymal stem cell surface marker CD44 and CD90 were detected positively using flow cytometry. CONCLUSION: A method for isolation,cultivation and preservation of human urine-derived stem cells was successfully established.

Key words: human urine-derived stem cells, primary culture, CD44, CD90

中图分类号:

Q813.1+1

周明, 孙震晓. 人尿源性干细胞的原代培养[J]. 癌变·畸变·突变, 2017, 29(5): 390-393.

ZHOU Ming, SUN Zhenxiao. Cultivation of human urine-derived stem cells[J]. Carcinogenesis, Teratogenesis & Mutagenesis, 2017, 29(5): 390-393.

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人尿源性干细胞的原代培养

Cultivation of human urine-derived stem cells

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