人胚胎干细胞向内皮细胞的分化诱导

发布时间：2018-05-06 21:45

本文选题：干细胞 + 胚胎干细胞　；参考：《中国组织工程研究》2016年23期

【摘要】：背景:人胚胎干细胞是一类特殊的细胞,具有自我更新及多向分化潜力,在一定诱导条件下能够分化为内皮细胞。目的:探讨人胚胎干细胞向内皮细胞分化的条件,研究血管内皮生长因子诱导人胚胎干细胞诱导分化为内皮细胞的效果。方法:取40代H9细胞进行复苏,采用悬浮培养法制备胚体,培养5 d后转入贴壁培养阶段,分化为拟胚体,添加含50μg/L血管内皮生长因子的胚胎干细胞培养基进行诱导培养。取诱导分化第2代细胞,进行DiI-Ac-LDL摄取实验;取诱导分化第15代细胞,进行免疫组织化学染色。结果与结论:(1)培养1 d时,可见条索或多角形的单层细胞在拟胚体细胞周围呈芽样放射状生长,细胞生长相对迅速,能够与周围集落的细胞相互融合;培养二三天后,悬浮细胞数得到进一步增加,但中心小圆形细胞开始死亡;培养5 d后,拟胚体细胞开始传代,细胞聚集呈现出典型铺路石样外观;(2)经过诱导分化后,部分人胚胎干细胞能够主动摄取荧光标记LDL,且细胞呈现为红色荧光颗粒;(3)免疫组化染色显示,诱导分化的第15代人胚胎干细胞呈CD31和FLK-1阳性;(4)结果表明:人胚胎干细胞在血管内皮生长因子诱导下可分化为内皮细胞。
[Abstract]:Background: human embryonic stem cells (human embryonic stem cells) are a special type of cells with potential for self renewal and multidirectional differentiation and can be differentiated into endothelial cells under certain induction conditions. Methods: the 40 generation H9 cells were recovered and the embryos were prepared by suspension culture method. After 5 d, the cells were transferred into the adherent culture stage and differentiated into embryoid bodies. The embryonic stem cell culture medium containing 50 mu g/L vascular endothelial growth factor was added to the culture medium for induction and differentiation of second generation cells, and DiI-Ac-LDL uptake experiments were carried out, and fifteenth generation cells were induced to be induced. Results and conclusions: (1) when the culture of 1 D, the monolayer cells of the cord or polygon showed a bud like growth around the embryoid cells, the cell growth was relatively rapid, and the cells could be fused with the surrounding colonies. After two or three days, the number of suspended cells was further increased, but the central small round cells were increased. After 5 d, the embryo somatic cells began to pass through the generation, and the cell aggregation showed a typical paved stone appearance. (2) after differentiation, some human embryonic stem cells could take the active uptake of fluorescent labeling LDL and the cells were red fluorescent particles; (3) the immuno histochemical staining showed that the differentiated fifteenth generation embryonic stem cells were CD31 and the induced differentiation. FLK-1 positive; (4) the results showed that human embryonic stem cells could be differentiated into endothelial cells induced by vascular endothelial growth factor.

【作者单位】：河南理工大学医学院;
【分类号】：R329.2

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