骨髓间充质干细胞分化为心肌细胞过程中Notch表达的研究

目的研究骨髓间充质干细胞分化为心肌细胞过程中Notch表达的研究。方法用密度梯度离心法分离培养犬骨髓间充质干细胞,按照酶法及差速贴壁法分离培养心肌细胞。观察干细胞增殖及传代情况。单独培养的干细胞为对照组,实验组将骨髓间充质干细胞与心肌细胞共培养,用RT-PCR、免疫细胞化学、MTT等方法检测干细胞分化为心肌细胞的情况,及干细胞在增殖与分化为心肌细胞过程中Notch信号系统的表达情况。结果骨髓间充质干细胞呈梭形、旋涡样生长,增殖及传代能力强,并可诱导分化为心肌样细胞,免疫荧光示心肌细胞标志物的表达。RT-PCR及免疫细胞化学显示实验组有Notch信号通路受体及配体的表达,而对照组表达微弱。结论骨髓间充质干细胞在增殖及分化过程中存在Notch信号通路,在干细胞分化为心肌细胞过程中Notch信号系统的表达上调。

The study of expression of Notch signaling in the differentiation of mesenchymal stem cells into cardiomyocytes

Objective To investigate the expression of Notch signaling of the differentiation of mesenchymal stem cells(MSCs) into cardiomyocytes.Methods MSCs were isolated from canine bone marrow and cultured with cell culture techniques,and then cardiomyocyte differentiation wad induced.The identification of the differentiation of MSCs and the Notch signal was carried out by RT-PCR and immunohistochemistry.MTT method was used to calculate the growth curve.Results MSCs looked fusiform under light microscopy.MSCs differentiated into cardiomyocytes.Immunofluorescence showed a significantly increased expression of TnT and CX43.RT-PCR and immunohistochemistry showed that the protein of the Notch signal was expressed during differentiation,and the level of expression was significantly lower during proliferation.Conclusion The Notch signaling system participates in the proliferation and differentiation of mesenchymal stem cells,and its expression increased during the differentiation of mesenchymal stem cells into cardiomyocytes.