摘要
目的探讨经血源子宫内膜间充质干细胞向子宫内膜上皮细胞分化的情况。方法选择2018年1月至2019年6月在新疆医科大学第一附属医院妇科中心体检的健康女性,收集经血并分离经血源子宫内膜间充质干细胞(menstrual blood-derived mesenchymal stem cells,MenSCs)进行培养,未加任何诱导药物的细胞作为对照组,分别在培养液中加入TGF-β110 ng/ml,EGF 10 ng/ml,PDGF-BB 10 ng/m1和与不同浓度的雌激素(17-β-雌二醇1×10^-7 mol/L、1×10^-8 mol/L、1×10^-9 mol/L),培养5 d后检测角蛋白(CK)和波形蛋白(VIM)蛋白与mRNA表达水平的变化,判断MenSCs是否向子宫内膜上皮细胞方向分化。结果经血分离原代细胞培养7 d后,长梭形纤维样细胞数目增多,细胞形成大克隆。随着传代次数的增加,细胞多呈现长梭形纤维样或纺锤状,MenSCs标记分子OCT-4表达率为(94.65±0.21)%、CD-45表达率为(1.86±0.19)%、Strol-1表达率为(94.65±0.21)%;CK mRNA在1×10^-9 mol/L组的表达水平显著高于对照组且显著低于1×10^-8 mol/L组(P<0.05),1×10^-7 mol/L组中CK mRNA表达水平显著高于对照组(P<0.05)、低于1×10^-8 mol/L组,但差异无统计学意义(P>0.05);VIM mRNA与对照组相比呈现剂量依赖性上调(P<0.05)。不同浓度17-β-雌二醇处理后MenSCs均呈现子宫内膜上皮细胞方向的分化。1×10^-7mol/L组中CK的荧光强度显著高于对照组细胞(P<0.05)。结论经血源子宫内膜间充质干细胞在适宜的条件下可诱导分化成子宫内膜上皮细胞,为妇产科疾病的研究提供了理想的实验材料与模型。
Objective To investigate the differentiation of endometrial mesenchymal stem cells derived from blood into endometrial epithelial cells.Methods From January 2018 to June 2019,healthy women who had physical examination in the Department of gynecology center of the First Affiliated Hospital of Xinjiang Medical University were selected to collect and separate the blood derived endometrial mesenchymal stem cells(menscs)for culture.The cells without any induction drugs were used as the control group,and TGF-β110 ng/ml,EGF 10 ng/ml,PDGF BB were added into the culture solution respectively 10 ng/ml and different concentrations of estrogen(17β-estradiol 1×10^-7 mol/L,1×10^-8 mol/L,1×10^-9 mol/L).After 5 days of culture,the expression levels of keratin(CK)and VIM protein and mRNA were detected to determine whether menscs differentiated to endometrial epithelial cells.Results After 7 days of primary culture,the number of long fusiform fibroid cells increased and the cells formed large clones.With the increase of subculture times,most of the cells showed long fusiform fiber or spindle shape,the expression rate of menscs marker Oct-4 was(94.65±0.21)%,the expression rate of cd-45 was(1.86±0.19)%,the expression rate of strol-1 was(94.65±0.21)%.The expression level of CK mRNA in 1×10^-9 mol/L group was significantly higher than that in the control group and significantly lower than that in 1×10^-8 mol/L Group(P<0.05),and the expression level of CK in 1×10^-7 mol/L group was significantly higher than that in the control group(P<0.05).The expression level of vim mRNA was significantly higher than that of the control group(P<0.05)and lower than that of the 1 x10-8 mol/L group,but there was no significant difference(P>0.05).After treatment with 17-β-estradiol at different concentrations,menscs showed differentiation in the direction of endometrial epithelial cells.The fluorescence intensity of CK in 1×10^-7 mol/L group was significantly higher than that in the control group(P<0.05).Conclusion Endometrial mesenchymal st
作者
赵蕾
陈志芳
杜蓉
袁琳
丁岩
ZHAO Lei;CHEN Zhifang;DU Rong;YUAN Lin;DING Yan(Center of Gynecology,First Af.liated Hospital of Xinjiang Medical University,Urumqi 830054,China)
出处
《中国妇产科临床杂志》
CSCD
北大核心
2020年第5期470-473,共4页
Chinese Journal of Clinical Obstetrics and Gynecology
基金
新疆维吾尔自治区自然科学基金资助项目(2016D01C280)。
