骨形态发生蛋白-2对大鼠脂肪干细胞成骨诱导的作用

发布时间：2018-04-11 11:06

本文选题：脂肪干细胞 + 骨形态发生蛋白-2　；参考：《中南大学》2009年硕士论文

【摘要】： 目的:探讨体外条件下不同浓度骨形态发生蛋白-2(BMP-2)对脂肪干细胞诱导分化为成骨细胞的影响。方法:本试验取3周龄雄性SD大鼠,颈部脱臼法处死,无菌条件下取出腹股沟脂肪垫。胶原酶-1消化分离,接种于含有胎牛血清的DMEM培养液中培养。在倒置显微镜观察细胞形态及生长情况。待细胞扩增并生长稳定后,分别加8组不同BMP-2浓度(根据每组BMP-2浓度不同,分为无BMP-2组、100ng/ml BMP-2组、200ng/mlBMP-2组、400ng/ml BMP-2组、800ng/ml BMP-2组、1000ng/mlBMP-2组、1200ng/ml BMP-2组、1600ng/ml BMP-2组)的诱导培养液进行成骨细胞诱导。用碱性磷酸酶、Ⅰ型胶原免疫组织化学染色及矿化结节染色鉴定成骨细胞分化能力。采用MTT比色法进行增殖分化活性比较。结果:碱性磷酸酶、Ⅰ型胶原免疫组织化学染色及矿化结节染色均为阳性。与对照组OD均值相比:200ng/ml BMP-2组、400ng/mlBMP-2组、800ng/ml BMP-2组、1200ng/ml BMP-2组、1600ng/mlBMP-2组有显著性差异(P＜0.05);100ng/ml BMP-2组、1000ng/mlBMP-2组无显著性差异(P＞0.05)。各组OD值均数随BMP-2浓度的增加先逐渐增大(0 ng/ml～400 ng/ml),达峰值后,又逐渐减少(800 ng/ml～1600 ng/ml)。结论:大鼠脂肪组织中分离培养出的脂肪干细胞(ADSCs)在体外具有向成骨细胞(OB)分化的潜能。当BMP-2浓度在400～800ng/ml时,能促进脂肪干细胞的增殖和分化。
[Abstract]:Aim: to investigate the effects of different concentrations of bone morphogenetic protein-2 (BMP-2) on differentiation of adipose stem cells into osteoblasts in vitro.Methods: male Sprague-Dawley rats aged 3 weeks were killed by cervical dislocations and inguinal fat pads were removed under aseptic condition.Collagenase-1 was digested and isolated and cultured in DMEM medium containing fetal bovine serum.Cell morphology and growth were observed under inverted microscope.The differentiation ability of osteoblasts was evaluated by alkaline phosphatase, type I collagen immunohistochemical staining and mineralized nodule staining.The activity of proliferation and differentiation was compared by MTT colorimetry.Results: alkaline phosphatase, type I collagen immunohistochemical staining and mineralized nodule staining were all positive.With the increase of BMP-2 concentration, the mean OD value of each group increased at first to 0 ng/ml~400 / ml ~ (-1), and then to the peak value, and then decreased to 800 ng/ml~1600 / ml ~ (-1) 路ml ~ (-1) 路L ~ (-1) 路L ~ (-1).Conclusion: adipose stem cells isolated from adipose tissue of rats have the potential to differentiate into osteoblasts in vitro.When the concentration of BMP-2 is 400~800ng/ml, it can promote the proliferation and differentiation of adipose stem cells.
【学位授予单位】：中南大学
【学位级别】：硕士
【学位授予年份】：2009
【分类号】：R329

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