摘要
目的探讨骨髓间充质干细胞(bone marrow stem cells,BMSCs)氯甲基苯甲酰胺(chloromethyl-benzamidodialkylcarbocyanine,CM-DiI)标记的效果及其标记后在大鼠阴道重建模型中的示综。方法全骨髓贴壁培养法培养大鼠BMSCs。用不同浓度的CM-Dil标记液标记BMSCs,观察不同浓度标记下BMSCs的标记效率和增殖能力。BMSCs与小肠黏膜下基质(small intestinal submucosa,SIS)复合后构建大鼠阴道重建模型,观察干细胞在重建阴道的示综。结果CM-DiI标记后各浓度的标记效率都可达98%以上,但荧光强度随浓度的增加而变强。2.5μmol/L及5μmol/L标记后的生长曲线与对照组无明显区别,10μmol/L及20μmol/L标记后生长曲线较对照组稍降低。BMSCs移植后主要分布在阴道深层组织中,术后3个月仍可见橙黄色的BMSCs。结论CM-DiI标记对干细胞活性影响小,5μmol/L可能是最佳的标记浓度,BMSCs移植后3个月仍可见橙黄色的BMSCs,可用于后期进一步研究。
Objective To explore the effect of chloromethyl-benzamidodialkylcarbocyanine(CM-Dil)labeling on bone marrow stromal stem cells(BMSCs)and its tracking in rat model of vaginal reconstruction.Methods Rat BMSCs were prepared with the whole bone marrow adherent method.BMSCs were labeled with different concentrations of CM-Dil and their labeling efficiency and reproductive activities were observed.The rat model of vaginal reconstruction was established with BMSCs seeded on small intestinal submucosal matrix(SIS).Its tracking in rat model of vaginal reconstruction was observed.Results BMSCs could be labeled with different concentrations of CM-Dil and the labeled efficiency was>98%,but the fluorescence intensity became stronger with the increase of concentration.The growth curves after labeling with the concentration of 2.5μmol/L and 5μmol/L were not significantly different from those of the control group,and the growth curves after labeling with 10μmol/L and 20μmol/L were slightly lower than those of the control group.After transplantation,BMSCs were mainly distributed in the deep vaginal tissues,and orange BMSCs could be found at 3 months after vaginal reconstruction.Conclusion CM-Dil did not affect stem cell activity,and 5μmol/L may be the optimal labeling concentration.Orange BMSCs could be found at 3 months after vaginal reconstruction and could be used in further study.
作者
张宁
秦锡静
朱玉林
李珮
黄向华
ZHANG Ning;QIN Xi-jing;ZHU Yu-lin;LI Pei;HUANG Xiang-hua(Department of Reproductive Medicine, the Fourth Hospital of Shijiazhuang City, Hebei Province, Shijiazhuang 050011, China;Department of Obstetrics and Gynecology, the Xiangya Changde Hospital, Hunan Province, Changde 415000, China;Department of Obstetrics and Gynecology, the Second Hospital of Hebei Medical University, Shijiazhuang 050000, China)
出处
《河北医科大学学报》
CAS
2021年第8期872-875,F0003,共5页
Journal of Hebei Medical University
基金
国家自然科学基金资助项目(81471418)
河北省医学科学研究重点课题计划(20201389)。
