人脐带间充质干细胞分离和初步鉴定

发布时间：2018-01-11 21:46

本文关键词：人脐带间充质干细胞分离和初步鉴定　出处：《中华实用诊断与治疗杂志》2016年06期 　论文类型：期刊论文

【摘要】：目的寻找一种高效获得稳定人脐带间充质干细胞的方法。方法采集3例足月剖宫产健康胎儿脐带,剥离血管及外膜得到Wharton’s胶。贴壁培养脐带Wharton’s胶,收集爬出的细胞进行传代,观察细胞形态;取第3代细胞采用流式细胞学检测细胞表面抗原的表达情况;使用成骨、成脂诱导分化的方法对第3代细胞进行分化潜能的鉴定,成骨诱导细胞用茜素红染液染色,成脂诱导细胞用油红O染液染色,并与未诱导分化的细胞进行比较。结果 Wharton’s胶培养10d左右可见梭形细胞从脐带组织逸出,贴壁培养细胞,可见呈放射状或螺旋状生长;流式细胞仪检测结果显示细胞表面抗原CD29、CD44呈高表达,CD34、CD45呈低表达;成脂诱导培养基中细胞油红O染色可见红色脂滴,成骨诱导培养基中细胞茜素红染色可见红色钙质结节,未诱导的细胞未见脂滴及结节样改变。结论贴壁培养的细胞符合间充质干细胞生长的形态,有向成骨、成脂分化的能力;贴壁法培养脐带Wharton胶可得到稳定的间充质干细胞。
[Abstract]:Objective to find an efficient stable human umbilical cord mesenchymal stem cells. Methods to collect 3 cases of full-term cesarean section healthy fetal umbilical cord, blood vessels and Wharton membrane stripping 's gel. The adherent culture of umbilical cord Wharton' s glue, climbed out of the collected cells were observed, cell morphology; the third generation the expression of cells by flow cytometry using cell surface antigen; osteogenic differentiation, identification method of adipogenic differentiation potential of the third generation cells, osteogenic cells were stained with alizarin red staining and adipogenic cells induced by oil red O dye staining, and differentiated cells were compared. The Wharton 's Glue 10d training around the spindle cells visible from the umbilical cord tissue escape of cultured adherent cells, visible radially or spiral growth; flow cytometry showed that the cell surface antigen CD29, CD44 showed high expression of CD34, CD45 expression was low; adipogenic cells cultured in medium oil red O staining showed red lipid droplets, osteogenic induction medium cells alizarin red staining showed red calcareous nodules, non induced cells were lipid droplets and nodular changes. Conclusion the adherent cells with mesenchymal stem cell growth morphology have osteogenic, adipogenic differentiation ability; cultured umbilical cord Wharton glue can be obtained by mesenchymal stem cells.

【作者单位】：郑州大学人民医院河南省人民医院妇产科;
【基金】：国家自然科学基金(81101685)
【分类号】：R329.2
【正文快照】： 间充质干细胞是干细胞家族中的一员,体外可大量扩增,具有分化成骨、软骨、脂肪的能力[1-2]。可用于多种疾病的治疗,如脊髓损伤、脑缺血、退变诱导的组织细胞变性等[3-5]。常见间充质干细胞来源有骨髓、脐血和脐带。骨髓间充质干细胞随年龄增加而减少,其分化能力也降低,取材时

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