摘要
本研究探讨健康人脐带间充质干细胞(human umbilical cord mesenchymal stem cells,hucMSC)中SDF-1受体CXCR4的表达特征。运用组织块贴壁法从健康人脐带中培养出hucMSC;采用流式细胞术检测hucMSC表面特异性标记;应用油红O染色及茜素红染色对hucMSC成脂和成骨情况进行检测;运用流式细胞术检测第2-第5代hucMSC中CXCR4受体的表达情况,同时采用实时荧光定量PCR法检测第2-第5代hucMSC中cxcr4mRNA的表达。结果表明,hucMSC表现出CD44、CD13、CD71阳性,CD38、CD117、HLA-DR阴性。hucMSC经成脂、成骨诱导后出现脂滴及钙结节,经油红O及茜素红染色均呈阳性。流式细胞术检测显示在第2-第5代hucMSC中均有CXCR4蛋白表达,分别为(89.82±0.62)%、(86.87±1.32)%、(80.50±4.46)%、(70.10±0.68)%。第2-第5代hucMSC中cxcr4mRNA均有表达,且第3-第5代cxcr4mRNA表达量分别为第2代的0.5585±0.0875倍、0.6205±0.1377倍、0.4634±0.0447倍。结论:从健康人脐带中培养的第2-第5代hucMSC均表达cxcr4mRNA,且其表达量随传代数增加而呈降低趋势;与第2代相比,第3-第5代hucMSC cxcr4mRNA表达均有所降低,但第3-第5各代间表达相对稳定。
The purpose of this study was to explore the expression characteristics of SDF-1 receptor,CXCR4, in mesenchymal stem cells（MSC） of different passages derived from human umbilical cord（hucMSC）.The hucMSC were isolated from Wharton′s jelly tissue of human umbilical cord by tissue culture.The expressions of specific marker in hucMSC were detected by flow cytometry.The adipogenic and osteogenic induction of hucMSC were detected by alizarin bordeaux and Oil red O staining.The expressions of CXCR4 protein in hucMSC of 2nd-5th passages were detected by flow cytometry,and cxcr4 mRNA levels in hucMSC of 2nd-5th passages were evaluated by real-time quantitative PCR.The results showed that the expression of CD44,CD13,CD71 were positive while CD38,CD117,HLA-DR were negative.After induced by osteogenic and adipogenic inductors,the lipid droplets and calcium nodals appeared in hucMSC,hucMSC stained with oil red O and alizarin red were shown to be positive.The cxcr4 was found in hucMSC of 2nd-5th passages,and their expressions were（89.82±0.62）%,（86.87±1.32）%,（80.50±4.46）%,（70.10±0.68）% respectively.The cxcr4 mRNA was found in hucMSC of 2nd-5th passages,and expression of cxcr4 of 3rd-5th passages were 0.5585±00875,0.6205±0.1377,0.4634±0.0447 times of expression of 2nd passage respectively.It is concluded that the cxcr4 mRNA expresses in hucMSC of 2nd-5th passages,and declines when the number of passages increases.Compared with 2nd passage,cxcr4 mRNA levels in hucMSC of 3rd-5th passages decline,but the expression level of cxcr4 mRNA between hucMSC of 3rd-5th passages is stable.
出处
《中国实验血液学杂志》
CAS
CSCD
2011年第1期169-173,共5页
Journal of Experimental Hematology
