不同氧分压时人脂肪干细胞细胞因子的分泌

Influence of different oxygen partial pressures on cytokines secreted from human adipose-derived stem cells

背景:不同氧分压对人脂肪来源干细胞分泌细胞因子的影响目前尚未定论,这些差异可能由于研究者对于氧分压的选取不同而造成影响。目的:检测不同氧分压对人脂肪来源干细胞分泌细胞因子的影响。方法:体外分离培养人脂肪来源干细胞进行免疫表型进行鉴定。将人脂肪来源干细胞分别在1%,3%,5%,10%,21%氧分压的环境中培养24 h后,使用实时定量PCR和酶联免疫吸附法对人脂肪来源干细胞分泌的血管内皮生长因子、肝细胞生长因子、神经细胞生长因子、角质细胞生长因子,在基因水平以及蛋白水平上进行检测分析。结果与结论:人脂肪来源的人脂肪来源干细胞阳性表达CD71,CD73,CD90,CD105,阴性表达CD34,CD45,CD54及HLA-DR。经单因素方差分析统计,在基因水平上,低氧环境(体积分数1%,3%氧)均可促进人脂肪来源干细胞显著性高表达血管内皮生长因子、神经生长因子(P均<0.01);对人脂肪来源干细胞表达肝细胞生长因子有显著影响(P<0.05);而对角质细胞生长因子的表达则无显著影响(P>0.05)。在蛋白水平上,低氧促进人脂肪来源干细胞分泌肝细胞生长因子、血管内皮生长因子蛋白(P均<0.01),对神经生长因子、角质细胞生长因子的分泌无显著影响。结果提示,人脂肪来源干细胞在低氧环境中培养后,其生物学特性受到一定的改变,可促进血管内皮生长因子、肝细胞生长因子、神经生长因子基因的表达,提高肝细胞生长因子、血管内皮生长因子蛋白的分泌。

BACKGROUND： Effects of different oxygen partial pressures on cytokine secretion of human adipose-derived stem cells have been differently reported. These differences may be caused by varying oxygen partial pressures. OBJECTIVE： To investigate the influence of different oxygen partial pressures on cytokines secreted from human adipose-derived stem cells.METHODS： Human adipose-derived stem cells were cultured in vitro and identified by its immunophenotype. Human adipose-derived stem cells were divided into five groups and cultured under different oxygen partial pressure conditions （1%, 3%, 5%, 10%, 21%）for 24 hours, respectively. With quantitative real-time PCR and enzyme linked immunosorbent assay, the secretion of cytokines, vascular endothelial growth factor, hepatocyte growth factor, nerve growth factor, keratinocyte growth factor, from human adipose-derived stem cells were analyzed on the gene and protein levels. RESULTS AND CONCLUSION： Human adipose-derived stem cells were positive for CD71, CD73, CD90, CD105 and negative for CD34, CD45, CD54, HI_A-DR. From the aspect of gene level, hypoxia （1%, 3% 02） promoted the expression of vascular endothelial growth factor and nerve growth factor from human adipose-derived stem cells （P 〈 0,01）, and significantly elevated the expression of hepatocyte growth factor （P 〈 0.05）; however, there was no significant influence on keratinocyte growth factor under hypoxia （P 〉 0.05）. Based on the protein level, protein secretion of hepatocyte growth factor and vascular endothelial growth factor from human adipose-derived stem ceils was increased under hypoxia （P 〈 0.01）, but no changes occurred in nerve growth factor and keratinocyte growth factor. After cultured under hypoxic environment, human adipose-derived stem cells were promoted to express gene vascular endothelial growth factor, hepatocyte growth factor and nerve growth factor, as well as to secrete protein keratinocyte growth factor and vascular endothelial growth factor.