摘要
目的采用改良组织块贴壁法制备人脐带间充质干细胞humanumbilicalcordmesenchymalstemcells，hUCMSCs）。方法无菌条件下取足月婴儿脐带，采用改良组织块贴壁法分离间充质干细胞，台盼蓝染色法计数单个核细胞数，倒置相差显微镜下观察细胞形态，流式细胞仪检测细胞周期及免疫表型。结果原代hUCMSCs的数量为（1-5）×10 5个。培养至8d，组织块边缘处有单细胞爬出；培养至2周，贴壁生长的细胞散在分布或形成小克隆；培养至20d，细胞呈梭型、三角形或多角形。培养第20天的原代hUCMSCs的G。／G，期细胞占88．12％，S期细胞占1．23％，G2／M期细胞占10．65％，大多数细胞处于细胞增殖的潜伏期，CD105阳性率为97．65％，CD34阳性率为2．54％。结论采用改良组织块贴壁法分离了hUCMSCs，其生物学特性与骨髓间充质干细胞相似，为其临床应用奠定了基础。
Objective To prepare human umbilical cord mesenchymal stem cells （hUCMSCs） by modified tissue ad- herence method. Methods The hUCMSCs were isolated from the umbilical cord of mature newborn aseptically by modified tissue adherence method, counted for mononuclear cells by trypan blue staining, observed for morphology under inverted phase contrast microscope, and tested for cell cycle and phenotype by flow cytometry. Results The count of primary hUCMSCs was （ 1 - 5 ） x 105. Single cells appeared at the edges of tissue 8 d after culture. Two weeks after culture, the adherent cells were distributed in scatter or formed small clones. However, 20 d after culture, the cells were in spindle, triangular or polygonal form. A portion of 88. 12% of primary hUCMSCs on day 20 after culture were at G0/Gl phase, while 1.23% at S phase and 10. 65% at G2/M phase. Most of the cells were in the incubation period of proliferation, of which 97.65% were positive for CD105, and 2. 54% for CD34. Conclusion The hUCMSCs were successfully isolated by modified tissue adherence method, of which the biological characters were similar to those of bone marrow mesenchumal stem cells. It laid a foundation of clinical application of hUCMSCs.
出处
《中国生物制品学杂志》
CAS
CSCD
2014年第3期415-418,共4页
Chinese Journal of Biologicals
基金
2012年内蒙古自治区区科技厅应用研发项目(20120402)
2013内蒙古医科大学英才培育项目(2013YCPY025)
关键词
脐带
间充质干细胞
组织块贴壁法
Umbilical cord
Mesenchymal stem cells
Tissue adherence method
