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目的研究人脐血间充质干细胞(MSCs)分离培养的生物学特性及其向成骨、成脂诱导分化的能力。方法从人脐血中分离扩增MSCs,显微镜下观察其形态及生长情况,绘制生长曲线,电镜下观察超微结构,流式细胞仪检测细胞表面标志物;成骨、成脂诱导后以碱性磷酸酶(ALP)染色、茜素红染色鉴定MSCs成骨分化潜能,油红O染色鉴定成脂分化潜能。结果人脐血MSCs为成纤维细胞样,漩涡状贴壁生长排列,传至第10代细胞形态无明显变化;电镜下显示为低分化细胞;细胞表面不表达CD34和CD45,强表达CD29、CD44和CD90;成骨诱导后可检测到ALP表达及钙化结节形成;成脂诱导后可检测到脂滴形成。结论人脐血中可分离出MSCs,与其他来源的MSCs具有类似的生物学特性及多向分化潜能,脐血有可能成为骨组织工程种子细胞的来源。
Objective To study the biological characteristics of human umbilical cord blood mesenchymal stem cells (MSCs) isolated and cultured and its ability to differentiate into osteoblasts and adipocytes. Methods MSCs were isolated and expanded from human umbilical cord blood. The morphology and growth of MSCs were observed under microscope. The growth curve was drawn. The ultrastructure was observed under electron microscope. The cell surface markers were detected by flow cytometry. After osteogenic and adipogenic induction, ALP staining and alizarin red staining were used to identify osteogenic differentiation potential of MSCs. Oil red O staining was used to identify adipogenic differentiation potential. Results Human umbilical cord blood MSCs were fibroblast-like and arranged in a swirling adherent growth pattern. There was no significant change in the morphology of passage 10 cells and low differentiated cells under electron microscope. The expression of CD34 and CD45 was not observed on the surface of cells, but strong expression of CD29 and CD44 And CD90. ALP expression and calcified nodules could be detected after osteogenic induction; lipid droplets could be detected after adipogenic induction. Conclusion MSCs can be isolated from human umbilical cord blood and have similar biological characteristics and multidirectional differentiation potential with other MSCs. Cord blood may be the source of bone tissue engineering seed cells.