无血清培养促进脂肪干细胞向血管内皮细胞分化

背景：无血清培养对大鼠脂肪干细胞向血管内皮细胞诱导分化影响的报道甚少。目的：观察无血清培养大鼠脂肪干细胞后向血管内皮细胞诱导分化的情况。方法：采用酶消贴壁培养法获得雄性SD大鼠脂肪干细胞,传代培养至第3代。实验组细胞无血清培养24 h,对照组用含体积分数10%胎牛血清的L-DMEM完全培养基培养。然后应用血管内皮细胞诱导培养基培养3周。结果与结论：大鼠脂肪干细胞经体外培养可呈多角形或梭形贴壁生长,并能稳定传代。传代后大鼠脂肪干细胞极低表达细胞表面标志CD31。大鼠脂肪干细胞定向血管内皮细胞诱导分化后细胞呈鹅卵石样,CD31阳性表达明显上升且实验组明显高于对照组。实验组诱导后大鼠脂肪干细胞能够吞噬Dil标记的乙酰化低密度脂蛋白及在基质胶上形成2D小管样结构,其能力明显强于对照组。结果证实无血清培养可促进体外大鼠脂肪干细胞向血管内皮细胞诱导分化。

Serum-free culture promotes rat adipose-derived stem cells differentiating into endothelial cells

BACKGROUND: There are few reports about the effect of serum-free culture on the differentiation of rat adipose-derived stem cells into vascular endothelial cells.OBJECTIVE: To investigate the isolation, serum-free culture of rat adipose-derived stem cells differentiating into vascular endothelial cells.METHODS:The rat adipose-derived stem cells were isolated from male Sprague-Dawley rats and expanded to the third passage by enzymatic digestion-adherent explants method. In the experimental group, rat adipose-derived stem cells were cultured in serum-free medium for 24 hours. In the control group, rat adipose-derived stem cells were cultured in low-glucose Dulbecco’s modified Eagle’s medium containing 10% fetal bovine serum. After that, the cells were cultured in inducing medium for 3 weeks to differentiate into vascular endothelial cells.RESULTS AND CONCLUSION: The rat adipose-derived stem cells grew as polygonal or fusiform-shaped adherent cells when cultured in vitro, which could stably proliferate and passage. The rat adipose-derived stem cells showed very low expression of CD31, a cell surface marker, after passages. After directional differentiations into vascular endothelial cells, the cells were pebble-shaped under the inverted microscope. Expression of CD31 was up-regulated, which was much higher in the experimental group than the control group. The induced cells in the experimental group had stronger abilities than those in the control group to swallow Dil-labeled acetylated low-density lipoprotein and form tube-like structures on the matrigel after differentiation into vascular endothelial cells. So, rat adipose-derived stem cells could be highly successfully induced to differentiate into vascular endothelial cells in vitro after serum-free culture.