催产素诱导脂肪来源细胞向心肌细胞方向分化

发布时间：2019-02-15 06:41

【摘要】：研究背景脂肪组织含有成熟脂肪细胞及脂肪组织来源干细胞(Adipose tissue derived stem cell,简称(?)ADSC)。成熟脂肪细胞在天花板培养下,吐脂变成具有干细胞特性的去分化脂肪细胞(Dedifferentiated FAT,简称DFAT)。ADSC和DFAT都已被证明具有分化为心肌样细胞的能力。催产素(Oxytocin,简称OT)被证明能将胚胎肿瘤干细胞P19及P19CL6诱导分化成心肌细胞,但它能否将ADSC及DFAT细胞诱导成心肌细胞尚无报道。实验目的检验OT能否将ADSC及DFAT诱导成为心肌样细胞。实验方法从脂肪中分离得到ADSC及成熟脂肪细胞。通过天花板培养足成熟脂肪细胞去分化为DFAT。按照不同诱导的时间(1周,2周,3周)及不同浓度(10-8M,10-7M,10-6M)分组对ADSC和DFAT进行诱导培养。通过RT-PCR,免疫荧光及Western Blot分别在基因水平和蛋白水平检测心肌特异性标记的表达。以人类心肌组织作为阳性对照,未加诱导的ADSC及DFAT细胞作为阴性对照。实验结果高浓度OT(10-6M)造成实验细胞大量死亡,无法进行后续试验。10-8M及10-7M浓度的催产素诱导三周后,ADSC和(?)DFAT在基因及蛋白水平检测到心脏特异性标记GATA4, NKX2.5,c-TnT的表达,但是未发现自主搏动现象。相同诱导条件下,两种细胞在基因和蛋白水平无显著差别。实验结论生理浓度和中等浓度的催产素可以将ADSC及DFAI诱导成为心肌样细胞。
[Abstract]:Background adipose tissue contains mature adipocytes and adipose tissue derived stem cells (Adipose tissue derived stem cell, (?) ADSC).) The ability of adipocytes to differentiate into cardiomyocyte-like cells (DFAT). ADSC and DFAT) has been proved to be able to differentiate into dedifferentiated adipocytes with stem cell characteristics in ceiling-cultured adipocytes. Oxytocin (Oxytocin,) has been proved to be able to induce the differentiation of embryonic tumor stem cells (P19 and P19CL6) into cardiomyocytes, but whether it can induce ADSC and DFAT cells into cardiomyocytes has not been reported. Objective to test whether OT can induce ADSC and DFAT into cardiomyocytes. Methods ADSC and mature adipocytes were isolated from fat. Dedifferentiation of mature adipocytes from ceilings to DFAT. ADSC and DFAT were cultured according to different induction time (1 week, 2 weeks, 3 weeks) and different concentration (10 -8 MN 10 -7 M 10 -6 M). The expression of myocardial specific markers was detected at gene level and protein level by RT-PCR, immunofluorescence and Western Blot, respectively. Human myocardial tissue was used as positive control and uninduced ADSC and DFAT cells as negative control. The results showed that high concentration of OT (10-6 M) resulted in a large number of cell death, which could not be followed up. After induction of oxytocin at 10-8 and 10-7 M concentrations for three weeks, The expression of cardiac specific marker GATA4, NKX2.5,c-TnT was detected in ADSC and (?) DFAT at gene and protein levels, but no autonomic pulsation was found. There was no significant difference in gene and protein levels between the two types of cells under the same induction conditions. Conclusion ADSC and DFAI can be induced into cardiomyocyte-like cells by oxytocin at physiological and medium concentrations.
【学位授予单位】：北京协和医学院
【学位级别】：硕士
【学位授予年份】：2011
【分类号】：R329

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