体外诱导脐带血管外周间充质干细胞向心肌细胞定向分化的初步研究  

Study on the differentiation of human umbilical cord perivascular cell into cardiac-like cells in vitro.

作　　者：胡煜[1] 佟铁壁[2] 程东凯[1] 宫立国[1] 阎影[3] 

机构地区：[1]沈阳东方医疗集团菁华医院,辽宁沈阳110006 [2]沈阳军区第二零二医院,辽宁沈阳110003 [3]美国密苏里大学医学院

出　　处：《中国实用内科杂志》2007年第S1期174-176,共3页Chinese Journal of Practical Internal Medicine

摘　　要：目的探讨应用5-氮胞苷(5-aza)诱导人脐带血管外周(HUCPV)细胞向心肌细胞分化的作用。方法足月剖宫产患者的脐带在无菌条件下用酶消化法收集原代 HUCPV 细胞,经体外纯化、扩增培养,取第3代细胞,用5×10^(-6)nmol/L 5-aza 诱导24 h 后继续培养4周,观察诱导分化结果。结果酶消化法可富集大量的HUCPV 细胞;诱导后相差显微镜下观察细胞呈长梭型排列,核浆比例明显降低,部分细胞胞浆内可见细丝样结构;免疫组织化学染色发现 HUCPV 细胞为 CD44^+、CD34^-的类骨髓间充质细胞,经诱导后均表达心肌特异性的α-肌动蛋白和肌钙蛋白 T;PCR 结果显示诱导后 a-cardiac action 和 cardiac troponin T 为阳性表达。结论HUCPV 细胞可作为细胞治疗的一个有效来源,经5-aza 诱导后可定向分化为幼稚心肌细胞。Objective To investigate the capability of cardiomyoganic differentiation of human umbilical cord perivascu- lar cells induced by 5-azacitidine in vitro.Methods Human umbilical cord perivascular colls were harvested from the umbilical cords of consenting full-term caesarian section patients,which were digested by collagenase,and were purified, expanded through repeating passaging in time in vitro.The HUCPV cell of passage 3 were treated with 5-azacytidine(5 -aza,5×10^(-6)nmol/L) for 24 hours and continued to be cultured under the previous condition for 4 weeks.The prolifera- tion and differentiated phenotype of HUCPV cells were observed.Results The isolation method of the UC vasculature and enzymatic digestion of its perivascular tissue to rapidly harvest a highly proliferative HUCPV cells.After treated with 5 -aza for 4 weeks,the passage 3 HUCPV cells formed the spindle-shaped cells and rate of nuclear and cytoplasm de- creased.Myotube-like structures were found in some cytoplasm.Immunohistochemistry results showed that the HUCPV cells were similar to MSC by stained positively for CD44 and negtively for CD34.The HUCPV cells induced by 5-aza were stained positively a-cardiac action and cardiac Troponin by immunohistochemistry.Furthermore,the expression of a -cardiac action and cardiac treponin gene as detected positively by RT-PCR after HUCPV cells were induced by 5- aza.Conclusion The HUCPV cell was a valuable material for cell-based therapies and can be induced into cardiac- like cells by treatment with 5-aza.

关 键 词：HUCPV 细胞 细胞分化 心肌细胞 

分 类 号：R329[医药卫生—人体解剖和组织胚胎学]