CM-DiI体外标记人乳牙牙髓干细胞的可行性研究

发布时间：2018-05-18 00:19

本文选题：人乳牙牙髓干细胞 + CM-DiI　；参考：《口腔医学研究》2016年03期

【摘要】：目的:筛选CM-DiI用于人乳牙牙髓干细胞标记的最佳浓度,并对标记后细胞的生物学行为进行评价,为下一步细胞活体移植和体内示踪打下基础。方法:酶解组织块法培养人乳牙牙髓细胞并纯化,免疫组化鉴定细胞来源,进行细胞体外多向诱导分化能力实验。将第3代细胞分别使用浓度2mg/L、3mg/L及4mg/L的CM-DiI进行标记,比较标记后细胞乳酸脱氢酶释放率及细胞增殖情况,并观察经体外多次传代后细胞荧光的表达情况。结果:酶解组织块法可获得成集落状生长的人乳牙牙髓细胞;免疫组化确定细胞为间充质而非造血来源;细胞具有成脂、成骨及成牙本质等多向分化能力。3种标记物浓度均未对细胞乳酸脱氢酶释放及细胞增殖产生显著的不利影响;随体外多次传代,标记荧光强度逐渐减退。结论:CM-DiI操作简便、染色速度快,可有效的标记人乳牙牙髓干细胞,其中3mg/L的标记浓度细胞毒性极小,标记效率高,在体外多次传代后荧光信号仍能稳定表达。
[Abstract]:Aim: to screen the optimal concentration of CM-DiI for labeling human deciduous dental pulp stem cells and evaluate the biological behavior of the labeled cells in order to lay a foundation for the next step of cell transplantation and in vivo tracer. Methods: human deciduous dental pulp cells were cultured and purified by enzymolysis tissue block method. The origin of cells was identified by immunohistochemistry and the ability of inducing differentiation in vitro was studied. The third passage cells were labeled with CM-DiI of 2 mg / L 3 mg / L and 4mg/L, respectively. The release rate of lactate dehydrogenase and cell proliferation were compared, and the fluorescence expression of the cells was observed after several passages in vitro. Results: colony forming human primary dental pulp cells could be obtained by enzymatic tissue block method. The cells were identified by immunohistochemistry as mesenchymal rather than hematopoietic sources. The concentration of 3 markers of osteogenesis and odontogenic ability had no significant adverse effect on the release of lactate dehydrogenase and cell proliferation, but the fluorescence intensity decreased gradually with the passage in vitro. Conclusion the 3mg/L labeled human dental pulp stem cells was easy to operate and stained quickly. The labeling concentration of 3mg/L was minimal and the labeling efficiency was high. The fluorescence signal could still be expressed stably after several passages in vitro.
【作者单位】：陕西省牙颌面疾病临床研究中心;西安交通大学口腔医院儿童牙病科;西安交通大学法医学院;
【基金】：陕西省科学技术研究发展计划项目(编号:2013K12-16-04)
【分类号】：R788

【相似文献】