论文部分内容阅读
为研究绵羊胚胎肌源性干细胞的生物学特性,取30日龄的绵羊胚胎,采用联合酶消化法及差速贴壁法分离培养获得均质的肌源性干细胞,并对其进行一系列生物学特性检测。结果显示,绵羊肌源性干细胞能传代培养至35代以上;免疫荧光染色和RT-PCR鉴定肌源性干细胞表面标记物Sca-1、CD34、CD144和Desmin呈阳性表达;生长曲线和细胞周期结果显示绵羊肌源性干细胞具有较强自我更新能力;通过不同的诱导方法可以将肌源性干细胞成功地诱导为肝样细胞和胰岛样细胞,Schiff染色和双硫腙染色均呈阳性,RT-PCR检测肝样细胞特异性标记物AFP和ALB以及胰岛样细胞特异性标记物Insulin均呈阳性表达,进一步证明了其诱导分化潜能。结果表明,本试验成功地分离培养获得了均质并具有较强自我更新潜能的绵羊肌源性干细胞,这种干细胞具有向肝样细胞和胰岛样细胞分化的潜能,这为临床治疗提供了潜在的理论基础和大量的种子细胞。
In order to study the biological characteristics of sheep embryonic myogenic stem cells, a 30-day-old sheep embryo was isolated and homogenized myogenic stem cells were isolated and cultured by a combination of enzyme digestion and differential adherent method. A series of biological Learning characteristics test. The results showed that sheep muscle-derived stem cells could be subcultured for more than 35 passages. The expression of Sca-1, CD34, CD144 and Desmin on myogenic stem cells was confirmed by immunofluorescence staining and RT-PCR. The growth curve and cell cycle The results showed that myogenic stem cells of sheep showed a strong self-renewal ability. Myogenic stem cells were successfully induced to hepatic-like cells and islet-like cells by different induction methods. Schiff staining and dithizone staining were all positive. RT-PCR Detection of hepatocyte-like markers AFP and ALB and islet-like cell-specific marker Insulin showed positive expression, further evidence of its potential for differentiation. The results showed that this experiment successfully isolated and cultured sheep muscle-derived stem cells that have a homogeneous and strong self-renewal potential and have the potential to differentiate into hepatocyte-like cells and islet-like cells, which provides potential for clinical treatment The theoretical basis and a large number of seed cells.