新生猕猴UCMSC的制备、鉴定及对老年猕猴自发DM的治疗作用

摘要

目的 探讨新生猕猴脐带间充质干细胞(UCMSC)对老年猕猴自发性糖尿病(DM)的治疗作用.方法 利用组织贴壁法获得猕猴UCMSC,流式检测UCMSC的免疫表型,进行体外分化能力分析.将UCMSC进行eGFP标记后,经静脉输入24周岁的自发DM猕猴体内.分别在UCMSC输入前和输入后1个月,测定DM猕猴血清葡萄糖(Glu)、甘油三酯(TG)、总胆固醇(TC)、血清胰岛素(INS)、血清C-肽(C-P)含量,比较输入前后的变化.结果 建立猕猴UCMSC分离、培养方法,并获得UCMSC,猕猴UCMSC高表达CD44和CD29,不表达CD34和CD45,可被诱导分化为软骨、骨、脂肪、胰岛样细胞.UCMSC输入到自发DM老年猕猴体内后,血清Glu、TG、TC水平降低,而INS、C-P水平升高.结论 成功分离扩增新生猕候UCMSC,该细胞对老年猕猴自发DM有一定治疗作用.%Objective To explore the therapeutic effect of umbilical cord-derived mesenchymal stem cells (UCMSCs) from newborn macaques on the spontaneous diabetes mellitus in senile macaques.Methods The tissue explants adherent method was conducted to obtain macaque USMSCs,flow cytometry was used to identify the immunophenotype of UCMSCs,and the in-vitro differentiation capability was analyzed.The UCMSCs were marked with eGFP and then given to 24 w-old macaques with spontaneous DM via intravenous infusion.One month before and after the UCMSC infusion,the levels of serum glucose (Glu),triglyceride (TG),total cholesterol (TC),serum insulin (INS) and serum C-peptide (C-P) were measured in macaques with DM respectively and the changes were compared.Results The isolation and cultivation method of macaque UCMSCs was established,and macaque UCMSCs were obtained;macaque UCMSCs expressed high levels of CD44 and CD29 but failed to express CD34 and CD45,and they could be induced to differentiate into chondrocytes,osteocytes,adipocytes and islet-like cells.After the UCMSC infusion to senile macaques with spontaneous DM,the levels of serum Glu,TG and TC reduced,while the levels of INS and C-P increased.Conclusion Newborn macaque UCMSCs are successfully isolated and proliferated;these UCMSCs have certain therapeutic effect on spontaneous DM in senile macaques.