成人脂肪干细胞的体外多向分化能力观察

发布时间：2018-05-21 13:42

  本文选题：成人脂肪干细胞 + 体外诱导　； 参考：《山东医药》2017年40期

【摘要】：目的观察成人脂肪干细胞(ADSCs)的体外多向分化能力。方法从正常成人脂肪组织分离得到ADSCs并体外培养。在ADSCs中分别加入成人脂肪、骨、软骨、肝细胞的体外诱导培养基诱导,观察诱导过程中细胞形态的变化,诱导前、诱导1周、诱导2周时采用RT-PCR法观察细胞相应标志物mRNA表达的变化。结果 ADSCs向脂肪细胞诱导2周,细胞内的脂质空泡聚集,小脂滴融合在一起并变大;最后细胞变为单房,并且充满脂质,红油O染色阳性;诱导1周细胞PPARγ、脂蛋白酶和αP2 mRNA表达较诱导前增加;诱导2周细胞PPARγ、脂蛋白酶mRNA表达降低,但αP2 mRNA变化不明显。ADSCs向骨细胞诱导5天,细胞由成纤维形转化为多边形或不规则状。诱导2周单层细胞聚集成块,形成细胞岛状结构,ALP染色阳性,von Kossa染色可见黑色矿化结节;矿化结节形成后细胞向结节移行聚集,促使更多的细胞形成更大的矿化结节;诱导后细胞ALP、骨桥蛋白和Ⅰ型胶原mRNA表达逐渐增加。ADSCs向软骨细胞诱导1周,细胞呈多边形或类圆形,细胞团向内呈放射状聚集,FITC染色后免疫荧光可见Ⅱ型胶原;诱导1、2周时细胞Ⅱ型胶原、聚集蛋白聚糖和SOX9 mRNA表达明显高于诱导前。ADSCs向肝细胞诱导2周时可见部分细胞变成圆形或椭圆形,并随时间顺延呈肝细胞样圆形细胞增多趋势,细胞核居中,圆形且边缘清楚,PAS染色阳性;诱导1、2周时细胞CK18、HNF1α及白蛋白mRNA表达明显高于诱导前。结论成人脂肪组织中分离得到的成人ADSCs可在体外定向分化为脂肪细胞、骨细胞、成骨细胞和肝细胞。
[Abstract]:Objective to observe the differentiation ability of adult adipose stem cells (ADSCs) in vitro. Methods ADSCs was isolated from normal adult adipose tissue and cultured in vitro. Adult fat, bone, cartilage and hepatocytes were added to ADSCs to induce in vitro. The changes of cell morphology were observed during the induction. The expression of mRNA was observed by RT-PCR method at 2 weeks after induction. Results after ADSCs was induced to adipocytes for 2 weeks, the lipid vacuoles gathered and the small lipid droplets fused together and became larger. Finally, the cells became monocyclic and filled with lipid, and the cells were positive for marked oil O staining. The expression of PPAR 纬, lipoprotein enzyme and 伪 P2 mRNA was increased at 1 week after induction, but the expression of PPAR 纬 and lipase mRNA decreased at 2 weeks after induction, but the change of 伪 P2 mRNA was not obvious. After 5 days of induction, the cells transformed from fibroblast to polygon or irregular. After 2 weeks, monolayer cells were induced to assemble into blocks to form the island structure of the cells, and black mineralized nodules could be seen by von Kossa staining, and after the formation of the nodules, the cells migrated to the nodules, which caused more cells to form larger mineralized nodules. After induction, the expression of ALP, osteopontin and type I collagen mRNA increased gradually. ADSCs were induced to chondrocytes for 1 week. The cells were polygonal or round. The expression of type 鈪，

本文编号：1919427