摘要
目的探究用细胞膜染料PKH26与DiR两种标记方法标记人脐带来源间充质干细胞(human umbilical cord mesenchymal stem cells,hUC-MSCs)在1型糖尿病小鼠(type 1 diabetes mellitus,T1DM)模型中的归巢情况,并分析两种示踪方法的优缺点。方法在体外分别用PKH26与DiR对细胞进行标记,观察其标记效果。将PKH26或DiR标记的hUC-MSCs通过尾静脉注射的方式移植入1型糖尿病小鼠体内,24h后观察hUC-MSCs在各脏器的分布情况。结果hUC-MSCs移植后24h,T1DM小鼠心脏、肝脏、脾脏、肺、肾脏、胰腺组织冷冻切片可观察到PKH26阳性的细胞。活体成像系统下可直观观察到DiR标记的hUC-MSCs在心脏、肝脏、脾脏、肺、肾脏、胰腺中的归巢。结论PKH26标记适用于荧光显微镜下观察受体组织切片中的hUC-MSCs,DiR标记适用于在活体成像系统中观察hUC-MSCs在活体动物组织中的分布,两种方法结合可从组织切片和整体上对hUC-MSCs进行示踪。
Objective To track PKH26 and DiR labeled human umbilical cord mesenchymal stem cells(hUC-MSCs)in T1DM mice after transplantation.Methods Frozen hUC-MSCs were resuscitated and labeled with PKH26 and DiR respectively.PKH26 or DiR labeled cells were intravenously injected into T1DM mice.Migration of PKH26 or DiR labeled hUC-MSCs were observed 24h after transplantation.Results PKH26 positive cells could be observed in heart,liver,spleen,lung,kidney and pancreas of T1DM mice after 24h.DiR positive signals could be detected in heart,liver,spleen,lung,kidney and pancreas of TIDM mice under small animal living imaging system.Conclusion PKH26 labeled cells is suitbale for being obeserved by microcopy in frozen sections.DiR labeled cells is suitbale for being obeserved by small animal living imaging system to know the distribution of hUC-MSCs.The combination of two methods could trace hUC-MSCs in vivo effectively.
作者
刘鹏
王从容
Liu Peng;Wang Congrong(Department of Endocrinology and Metabolism,Shanghai Jiaotong University Affiliated Sixth People′s Hospital,Shanghai 200233,China)
出处
《医学研究杂志》
2021年第11期55-59,共5页
Journal of Medical Research
基金
国家自然科学基金资助项目(82070913)
上海市科学技术委员会基金资助项目(20ZR1446000)。
