地黄低聚糖对体外培养的小型猪脂肪组织来源干细胞增殖的影响

Effect of Rehmannia glutinosa oligosaccharides on proliferation of miniswine adipose tissue-derived stem cells in vitro

目的分离培养小型猪脂肪组织来源干细胞(ASCs),观察不同浓度地黄低聚糖(RGOs)对其增殖的影响。方法酶消化法及贴壁法从小型猪腹股沟脂肪组织中分离、培养ASCs,流式细胞仪检测ASCs表面抗原CD29、CD31、CD34、CD44、CD45、CD90、CD105、HLA-DR的表达,CCK-8比色法观察不同浓度RGOs(0g/L、0.001g/L、0.01g/L、0.1g/L、1g/L、10g/L)对其增殖的影响。结果第4代ASCs表面抗原CD29、CD44、CD90、CD105表达呈阳性,CD31、CD34、CD45、HLA-DR表达呈阴性。RGOs在0.01～1 g/L浓度范围内对体外培养的小型猪ASCs具有促增殖作用(P<0.05),最佳浓度为0.1 g/L。结论一定浓度的RGOs对体外培养的小型猪ASCs具有促增殖作用。

Objective To isolate and culture miniswine adipose tissue-derived stem cells （ASCs） in vitro and investigate the effect of Rehmannia glutinosa oligosaceharides （RGOs） on its proliferation. Methods The isola- tion and culture of miniswine ASCs was conducted by enzyme digestion and adherent from adipose tissue harvested from inguinal regions. The surface antigens （ CD29, CD31, CD34, CD44, CD45, CD90, CD105, HLA-DR） of ASCs were examined by flow cytometry,and the proliferative effect of RGOs （concentration：0 g/L,0.001 g/L,0.01 g/L, 0.1 g/L,1 g/L and 10 g/L） was observed by CCK-8 colorimetry. Results The ASCs of the forth passage was posi- tive for CD29 ,CD44 ,CD90 and CD105 ,but negative for CD31 ,CD34 ,CD45 and HLA-DR. According to CCK-8 color- imetry result,RGOs accelerated proliferation of ASCs in vitro in the concentration ranged from 0.01 g/L to 1 g/L （ P 〈 0. 05 ）, and the best proliferative effect was observed at the concentration of 0. 1 g/L. Conclusion RGOs could accelerate proliferation of ASCs in vitro.