Wnt3a诱导小鼠胚胎干细胞向心肌细胞分化的研究

目的：研究Wnt3a在诱导小鼠胚胎干细胞心肌细胞分化中的作用和原理。方法：设计不同浓度，不同成分的Wnt3a条件培养基对小鼠胚胎干细胞诱导分化，对分化细胞进行形态学鉴定，通过免疫细胞化学检测心肌肌钙蛋白-T（cTnT）的表达，通过RT．PCR检测肌球蛋白重链（ot．MHC）和肌球蛋白轻链（MLC．2v）的表达。结果：Wnt3a诱导小鼠胚胎干细胞分化为心肌样细胞，分化细胞具有自动收缩性，免疫细胞化学检测心肌肌钙蛋白．T（cTllT）表达阳性，RT．PCR检测肌球蛋白重链（d—MHC）和肌球蛋白轻链（MLC-2v）表达阳性。经典Wnt信号途径的抑制剂Frizzled一8／Fc，能够抑制Wnt3a的诱导分化作用。结论：Wnt3a通过经典Wnt信号途径诱导小鼠胚胎干细胞向心肌细胞分化。

Effect of Wnt3a in Inducement of Mouse Embryonic Stem Cells to Differentiate into Cardiac Muscle Cell

Objective： To investigate the role and principle of Wnt3a in the induction of mouse embryonic stem ceils cardiac differentiation. Methods： Different concentrations and components of Wnt3a conditioned medium were used to induce mouse ES cells. Differentiated cells were identified on the morphology. Cardiac troponin-T （cTnT） is detected by immunocytochemieal, Myosin heavy chain （~-MHC） and myosin light chain（MLC -2v） is detected by RT-PCR. Results： Wnt3a induces mouse ES cells to differentiate into myocardial cells that contraction is automatic. Cardiac troponin-T （cTnT） expression is positive by immunocytochemical. Myosin heavy chain （~-MHC） and myosin light chain （MLC -2v） expresses positive by RT-PCR. Canonical Wnt signaling pathway inhibitors Frizzled-8/Fc could inhibit the differentiation efficiency of Wnt3a. Conclusion： Wnt3a induces cardiac differentiation of embryonic stem cells through canonical Wnt signal pathway.