SHE Tao, HU Da-hai, ZHANG Jun, LIU Jia-qi, ZHANG Wan-fu, CAI Wei-xia, ZHAO Zhou-ting, TANG Chao-wu
Burn Center of PLA, Xijing Hospital, Fourth Military Medical University, Xi'an 710032, China.
Zhonghua Shao Shang Za Zhi. 2009 Aug;25(4):268-71.
To study the effect of insulin in different concentrations on secretion function of growth factors of adipose-derived stem cells (ADSCs).
ADSCs were isolated from human abdominal adipose tissue and cultured. The immunophenotype and adipose induced-differentiation were identified, and the third generation cells were collected. The collected cells were assigned to 1 x 10(-8), 1 x 10(-7), 1 x 10(-6) mol/L insulin groups according to the concentration of added insulin. When cells grew into 70% confluence in conventional medium, ADSCs were cultured further in serum-free DMEM containing insulin in different concentrations for 3 days. ADSCs cultured in medium without insulin were used as control group. Secretion amount of vascular endothelial growth factor (VEGF) and hepatocyte growth factor (HGF) of ADSCs were determined by enzyme-linked immunosorbent assay. The effects of the supernatant fluid of ADSCs' nutrient solution on the proliferation and collagen synthesis of the cultured fibroblast were detected by MTT chromatometry and hydroxyproline chromatometry.
The secretion amounts of VEGF and HGF of ADSCs in 1 x 10(-8) and 1 x 10(-7) mol/L insulin groups [(471 +/- 41, 762 +/- 66 ng/L), (643 +/- 64, 930 +/- 67 ng/L), respectively] were significantly higher as compared with those in control group (286 +/- 47, 577 +/- 84 ng/L) (P < 0.05 or P < 0.01). No change occurred in the secretion amount of VEGF and HGF of ADSCs in 1 x l0(-6) mol/L insulin group (P > 0.05). The supernatant fluid of ADSCs' nutrient medium of 1 x 10(-8), 1 x 10(-7) mol/L insulin groups showed obvious stimulative effect on the proliferation and collagen synthesis of fibroblasts, and it was most obvious in the 1 x 10(-7) mol/L group (P < 0.05 or P < 0.01).
Insulin in the concentrations of 1 x 10(-8) and 1 x 10(-7) mol/L can notably promote ADSCs' function of secreting VEGF and HGF.
研究不同浓度胰岛素对脂肪干细胞（ADSCs）生长因子分泌功能的影响。
从人腹部脂肪组织中分离并培养ADSCs。鉴定其免疫表型及脂肪诱导分化情况，收集第三代细胞。根据添加胰岛素的浓度将收集的细胞分为1×10⁻⁸、1×10⁻⁷、1×10⁻⁶mol/L胰岛素组。当细胞在常规培养基中生长至70%汇合时，将ADSCs在含不同浓度胰岛素的无血清DMEM中进一步培养3天。将在无胰岛素培养基中培养的ADSCs作为对照组。采用酶联免疫吸附测定法测定ADSCs中血管内皮生长因子（VEGF）和肝细胞生长因子（HGF）的分泌量。通过MTT比色法和羟脯氨酸比色法检测ADSCs培养液上清对培养的成纤维细胞增殖和胶原合成的影响。
1×10⁻⁸和1×10⁻⁷mol/L胰岛素组ADSCs的VEGF和HGF分泌量[分别为（471±41，762±66 ng/L），（643±64，930±67 ng/L）]显著高于对照组（286±47，577±84 ng/L）（P<0.05或P<0.01）。1×10⁻⁶mol/L胰岛素组ADSCs的VEGF和HGF分泌量无变化（P>0.05）。1×10⁻⁸、1×10⁻⁷mol/L胰岛素组ADSCs培养液上清对成纤维细胞的增殖和胶原合成有明显促进作用，在1×10⁻⁷mol/L组最明显（P<0.05或P<0.01）。
1×10⁻⁸和1×10⁻⁷mol/L浓度的胰岛素可显著促进ADSCs分泌VEGF和HGF的功能。
