摘要
目的利用多能干细胞与仿生材料支架构建人工心肌,探讨仿生材料支架本身对多能干细胞心肌特异性分化的直接作用,为构建组织工程心肌提供理论支持。方法采用静电纺丝聚己内酯纳米纤维仿生支架,接种小鼠i PSCs(mi PSCs)细胞培养分化15 d,免疫荧光染色与Western blot法检测多能干细胞与心肌细胞特异性标志物。结果mi PSCs特异性表达多能干性标志物Oct4和Nanog,而且能够在聚己内酯纳米纤维支架上集落样增殖、分化;培养15 d后,mi PSCs在支架上分化出心肌特异性标志物c Tn T和MLC2a双重免疫荧光染色阳性的细胞;心肌细胞特异性结构蛋白c Tn T、α-MHC和MLC2的表达水平,支架组全面高于对照组。结论聚己内酯纳米纤维仿生支架支持mi PSCs生长与心肌细胞特异性分化。
Objective To explore the possible influence of biomimetic scaffolds on cardiomyocyte（ CM） differentiation of induced pluripotent stem cells（ i PSCs）. Methods The Oct4-GFP＋mouse i PSCs（ mi PSCs） were seeded directly on the poly-（ ε-caprolactone）（ PCL） nanofibrous biomimetic scaffolds,which were fabricated by an electrospun technology,and differentiated for 15 days. The development of CMs was verified by both immunofluorescence staining and Western blot detection. Results The results showed that mi PSCs,expressing Oct4 and Nanog before differentiation,proliferated on the PCL nanofibrous scaffold and grew in colonies during spontaneous differentiation period. Immunofluorescence staining showed the mi PSCs-derived cells were co-labeled with cardiomyocyte specific proteins,such as cardiac Troponin T（ c Tn T） and myosin light chain 2 a（ MLC2 a）. Moreover,Western blot demonstrated that cells derived from mi PSCs after differentiation for 1 5 days expressed higher level of c Tn T,MLC2 and α-myosin heavy chain（ α-MHC） than controls. Conclusions Theses results suggest that the PCL nanofibrous scaffold may support proliferation and cardiomyocyte differentiation of the Oct4-GFP＋mi PSC.
出处
《基础医学与临床》
CSCD
2016年第4期486-491,共6页
Basic and Clinical Medicine
基金
国家自然科学基金(31271039
31400832)
关键词
诱导多能干细胞
聚己内酯
纳米纤维支架
心肌细胞
induced pluripotent stem cell
poly-（ε-caprolactone）
nanofibrous scaffold
cardiomyocyte
