摘要
目的分离兔脂肪基质细胞并诱导培养其成骨表型,为扩增种子细胞提供实验依据。方法获取兔脂肪组织,胶原酶消化得到脂肪基质细胞,进行原代培养,消化传代后诱导培养,设置含重组人骨形态发生蛋白7(rhBMP-7)、转化生长因子β1(TGF-β1)的改良成骨诱导化培养组、常规成骨诱导培养组及对照非成骨诱导培养组,绘制生长曲线并对其成骨表型进行鉴定。结果与常规成骨诱导化方式相比,rhBMP-7、TGF-β1能明显促进的脂肪基质细胞增殖,并促使其向成骨细胞演变,碱性磷酸酶及VonKossa染色强阳性,群体倍增时间为32h;对照组未显示成骨细胞方向分化。结论采用多因子的成骨诱导培养利于脂肪基质细胞的增殖及诱导分化,是扩增组织工程种子细胞的有效方法。
Objective To supply an experimental data for augmentation of seed cells by culturing adipose derived stromal cells in osteoinductive manner. Methods Primary culture and passage culture of adipose derived stromal cells were made by method of collagenase digestion. Modified osteoinductivc culture group containing rhBMP - 7 and TGF - β1, routine osteoinductive and control culture groups were established. The cells were observed under inverted microscope daily and cells were assayed with growth curve. Meanwhile, osteogenic phenotype was determined by alkaline phosphate and Von Kossa staining. Results Compared with routine oeseoinductive culture group, rhBMP - 7 and TGF - β1 could stimulate the proliferation of adipose derived stromal cells and also promoted the osteogenic differentiation of cultured cells. The population doubling time was 32 hours. ALP and Von Kossa staining displayed significant positive expression and formation of mineralized nods in extracellular matrix. While, there was no osteogenic tendency in control group. Conclusion Multiple growth factor based osteoindcutive culturing is able to stimulate the proliferation and osteogenic differentiation of adipose derived stromal cells, which acts as an effective way to augment seed cells for tissue engineering.
出处
《中国骨与关节损伤杂志》
2007年第11期922-924,共3页
Chinese Journal of Bone and Joint Injury
基金
天津市卫生局课题基金资助项目(06KZ46)
