血管内皮细胞和脂肪干细胞体外联合培养细胞的增殖

Proliferation of vascular endothelial cells and adipose tissue-derived stem cells under in vitro coculture

背景:血管内皮细胞可有力支持脂肪干细胞向成骨方向转变,还可以加快血管的发生,为干细胞成骨提供营养支持。目的:探讨血管内皮细胞和脂肪干细胞体外联合培养对混合细胞增殖的影响。方法:设立单纯血管内皮细胞培养组、脂肪干细胞与血管内皮细胞按3∶1,1∶3,1∶1混合培养组、单纯脂肪干细胞培养组,加入含胎牛血清的L-DMEM,置于37℃、体积分数为5%的CO2饱和湿度培养箱中。倒置显微镜下观察细胞形态变化,改良MTT法绘制细胞生长曲线。结果与结论:脂肪干细胞与血管内皮细胞按3∶1,1∶1混合培养14d,细胞之间出现许多突触连接,部分细胞融合成团块状,其余培养方式均未见细胞团出现。随培养时间的延长,各培养方式下细胞吸光度值均逐渐升高,单纯脂肪干细胞培养、脂肪干细胞与血管内皮细胞按3∶1,1∶3,1∶1混合培养12d时细胞吸光度值达高峰,且按1∶1混合培养的吸光度值最高;而单纯血管内皮细胞培养10d吸光度值达高峰;随后各培养方式下细胞吸光度值逐渐下降。提示血管内皮细胞与脂肪干细胞体外联合培养,细胞能够相互促进增殖,1∶1混合比例条件下细胞增殖能力最强。

BACKGROUND：Vascular endothelial cells （VECs） have the ability to enhance adipose tissue-derived stem cells （ADSCs） to ossify and new blood vessel can provide nutrition for the ossification of stem cells. OBJECTIVE：To explore the influence of coculture of VECs and ADSCs on cell proliferation in vitro. METHODS：We set simple VECs group,3：1,1：3,1：1 coculture group and simple ADSCs group. Cells were incubated in L-DMEM containing fetal bovine serum at 37 ℃ in a 5% CO2 saturated humidity incubator. Morphological changes were observed by inverted microscope. Cell growth curve was drawn using modified MTT assay. RESULTS AND CONCLUSION：ADSCs and VECs were mixed at 3：1 and 1：1 for 14 days. Many synaptic linkages were visible among cells. Partial cells fused into a mass. Cell masses were not observed by other culture methods. With prolonged time,the absorbance of every group was increased gradually and reached a peak in ADSCs,3：1,1：3 and 1：1 ratio groups on day 12 and the highest was the 1：1 group,but the time of reaching the peak was on day 10 in the VECs group. The absorbance was gradually reduced under other culture methods. Results have indicated that VECs and ADSCs can stimulate proliferation each other in the system of co-culture in vitro; it can reach the best result when the proportion is 1：1.