碱性成纤维细胞生长因子对人前脂肪细胞增殖和分化的影响

背景：以前的观点认为碱性成纤维细胞生长因子通过促进移植脂肪内的血管再生来提高脂肪的存活率，但其对脂肪细胞的影响没有明确的定论。 目的：观察碱性成纤维细胞生长因子对人前脂肪细胞增殖和诱导分化的影响。 设计、时间及地点：细胞学体外培养，对比观察，于2006-03/2007-12在武警医学院完成。 材料：获取腹部吸脂术患者的脂肪组织标本10例，男1例，女9例，年龄21~45岁。 方法：通过分离、培养和自然纯化过程，获取人前脂肪细胞。实验随机分为碱性成纤维细胞生长因子组和对照组，对照组细胞单纯应用DMEM培养基进行培养传代，碱性成纤维细胞生长因子组使用稀释质量浓度为100 ng/L的碱性成纤维细胞生长因子DMEM培养基对细胞生长进行干预。 主要观察指标：①倒置相差显微镜观察培养后0，3，6，9，12，15 d细胞形态，并进行细胞计数，测定细胞生长曲线。②应用油红O染色，观察细胞内脂肪聚集情况，酶联免疫检测仪上测定吸光度值。 结果：接种后第1~15天，可见梭形的前脂肪细胞分裂增殖，局部融合成细胞单层，部分细胞分化，细胞内脂滴聚集增多。已有细胞变为单泡细胞，接近发育成熟。碱性成纤维细胞生长因子组和对照组细胞形态无明显差异，但碱性成纤维细胞生长因子组细胞数比对照组细胞数多44%。油红O染色后人前脂肪细胞内逐渐出现脂滴，呈暗红色，在培养后15 d细胞内脂滴浓度达到顶峰。碱性成纤维细胞生长因子组吸光度值比对照组增加300%，差异具有显著性意义(P < 0.05)。 结论：碱性成纤维细胞生长因子可以促进人前脂肪细胞的增殖及向脂肪细胞分化。

Effect of basic fibroblast growth factor on the proliferation and differentiation of human preadipocytes

BACKGROUND: Previous studies have shown that basic fibroblast growth factor (bFGF) can enhance fat survival rate by promoting vessel regeneration of transplanted fat. However, there is no clear conclusion addressing its effects on adipocytes. OBJECTIVE: To investigate the effect of bFGF on the proliferation and differentiation of human preadipocytes. DESIGN, TIME AND SETTING: Cytological in vitro comparison observation was conducted at the Medical College of Chinese People s Armed Police Force from March 2006 to December 2007. MATERIALS: Totally 10 samples of adipose tissue from patients undergoing abdominal liposuction were used in this study, including 1 male and 9 females, aged 21-45 years. METHODS: The human preadipocytes were obtained from human abdominal fat particles by isolation, culture and natural purity. The experiment was divided into bFGF and control groups. Cells in the control group were incubated in DMEM. Cells in the bFGF group were incubated in DMEM supplemented with 100 ng/L bFGF. MAIN OUTCOME MEASURES: Morphology of cultured cells was studied to determine the growth curve at 0, 3, 6, 9, 12, 15 days following culture by inverted phase contrast microscope. Fat aggregation was observed in cells using oil red staining. Absorbance value was examined using enzyme-linked immunosorbent assay. RESULTS: The preadipocytes represented the spindle-like character with strong proliferation. During 1 to 15 days, the preadipocytes proliferated into monolayers. Some cells became single bubble cells with lipid droplets deposited, close to maturity. The cells in bFGF group had no significant differences with the control group in cell morphology, but the amount of cells in bFGF group was 44% more than the control group. Following oil red O staining, lipid droplet was found in the preadipocytes, showing dark red. Following 15 days of incubation, the concentration of lipid droplet reached a peak. The absorbance of bFGF group was increased 300% compared with the control group (P < 0.05). CONCLUSION: The bFGF not only promote the proliferation of human preadipocytes, but also induce the differentiation of preadipocytes into mature adipocytes.