大鼠脂肪干细胞分离培养及细胞表型的研究

发布时间：2018-04-09 08:09

本文选题：脂肪干细胞　切入点：细胞培养　出处：《海南医学》2014年09期

【摘要】：目的分离、培养和冻存大鼠脂肪干细胞,并对其相关的细胞表型进行研究,为利用脂肪干细胞治疗梗阻性肾病肾纤维化等实验提供依据。方法取大鼠3只,经消毒麻醉,采集其腹股沟处脂肪组织分离培养其中的脂肪干细胞,用相差倒置显微镜观察细胞生长方式及形态变化。取第三代细胞用流式细胞仪作细胞免疫表型的鉴定。冻存复苏后再次检测干细胞生长情况及表型。结果大鼠脂肪组织中含有大量间充质干细胞,原代培养的ASCs呈小圆形或星形,经传代后的脂肪干细胞形态比较均一,呈长梭形,个别略呈多角形、漩涡样生长。其细胞表型为CD29、CD44高表达,CD73、CD105中等程度表达,CD34极低表达。脂肪干细胞经低温冻存3个月,复苏后,生长活力和存活率与冻存前未见明显区别,CD44和CD29检测与冻存前表达率也一致。结论建立了一种脂肪干细胞的有效分离、培养及保存方法,并对其表面标志物进行鉴定,为利用脂肪干细胞进行进一步的实验研究提供了依据。
[Abstract]:Objective to isolate, culture and freeze adipose stem cells from rats, and to study the phenotype of adipose stem cells in order to provide evidence for the treatment of renal fibrosis with obstructive nephropathy by adipose stem cells.Methods the adipose stem cells were isolated from adipose tissue in groin of 3 rats after disinfectant anesthesia. The growth patterns and morphological changes of the cells were observed by phase contrast inverted microscope.The third generation cells were identified by flow cytometry.After cryopreservation and resuscitation, the growth and phenotype of stem cells were examined again.Results there were a large number of mesenchymal stem cells in adipose tissue of rats. The primary cultured ASCs was small round or star-shaped. After passage, the adipose stem cells were uniform in shape and long fusiform, and some of them were polygonal and whirlpool.The phenotype of CD29 and CD44 was high and CD73 and CD105 were moderately expressed and the expression of CD34 was extremely low.After cryopreservation for 3 months, the growth activity and survival rate of adipose stem cells were not significantly different from those before cryopreservation and the expression rates of CD44 and CD29 were the same.Conclusion an effective method for the isolation, culture and preservation of adipose stem cells was established, and its surface markers were identified, which provided a basis for further experimental study on adipose stem cells.
【作者单位】：成都市第五人民医院病理科;成都中医药大学附属医院检验科;成都中医药大学附属医院中心实验室;
【基金】：四川省卫生厅资助项目(编号:110520) 成都市科技局资助项目(编号:313-106) 成都中医药大学附属医院基金项目(编号:Y2009003)
【分类号】：R329.2

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