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目的:研究人脐带间充质干细胞(UCMSCs)多向分化潜能,了解UCMSCs移植治疗脑缺血缺氧损伤(HIBD)大鼠的神经修复功能。方法:采用组织消化原代贴壁法分离培养人UCMSCs;取第4和10代UCMSCs进行流式细胞术检测间充质干细胞(MSCs)表面特异标志物表达;染色体分型检测染色体是否畸变;进行成神经、骨、脂诱导,了解其多向分化潜能;制备HIBD大鼠模型,穿梭箱测试对照组、HIBD细胞移植组和HIBD组大鼠的主动逃避率(AARR)和未逃避率(NARR)。结果:流式细胞术结果显示,类似MSCs样细胞强表达CD29、CD44和CD105,极低表达CD34和CD45,符合UCMSCs的特征。多向分化诱导后的细胞染色和免疫荧光结果显示,UCMSCs可诱导分化为神经样细胞、脂肪细胞和成骨细胞,多次传代后仍可多向诱导,且未发生染色体畸变。穿梭箱测试提示,UCMSCs移植组大鼠在测试的第3、4和5天的AARR高于HIBD组(P<0.05)。结论:UCMSCs易获取及纯化,具有多向分化潜能,多次传代功能稳定,可改善脑损伤动物的神经功能。
OBJECTIVE: To investigate the multidirectional differentiation potential of human umbilical cord mesenchymal stem cells (UCMSCs), and to understand the neurological repair function of UCMSCs transplantation in rats with cerebral ischemia-hypoxia injury (HIBD). Methods: Human UCMSCs were isolated and cultured by primary digestion method. UCMSCs of passage 4 and passage 10 were used to detect the expression of specific surface markers of mesenchymal stem cells (MSCs) by flow cytometry. Chromosome typing was used to detect whether the chromosomes were abnormal. The AARR and NARR of rats in HIBD rat model, shuttle box test control group, HIBD cell transplantation group and HIBD group were determined. . Results: The results of flow cytometry showed that MSCs-like cells strongly expressed CD29, CD44 and CD105, and very low expression of CD34 and CD45, which was in accordance with the characteristics of UCMSCs. Cell differentiation and immunofluorescence after induction of differentiation showed that UCMSCs could differentiate into neuron-like cells, adipocytes and osteoblasts. After many passages, UCMSCs could be induced more directionally without chromosome aberration. The shuttle box test showed that the AARR of UCMSCs transplantation group was higher than that of HIBD group on the 3rd, 4th and 5th day of the test (P <0.05). CONCLUSION: UCMSCs are easy to acquire and purify, have multidirectional differentiation potential, and have multiple functions of passage and stability, which can improve the neurological function of brain injured animals.