【摘要】 目的 :分离人体脂肪基质细胞并诱导培养其成骨表型。方法 :将脂肪抽吸术获取的人体脂肪进行机械分割 ,通过Ⅰ型胶原酶消化后得到脂肪基质细胞 ,在BGJb 培养基中原代培养 10d ,消化传代后用含体积分数 10 %FBS及 10 - 8mol/L地塞米松 ,50mg/L左旋抗坏血酸 ,10nmol/L维生素D3 ,10mmol/Lβ 磷酸甘油钠的DMEM /F12培养基中诱导培养 14d ,观察细胞形态 ,绘制细胞生长曲线并对其成骨表型进行鉴定。结果 :脂肪基质细胞呈成纤维细胞样贴壁生长 ,其中的成体干细胞经诱导培养后体积明显增大 ,胞核大面圆 ,胞浆丰富 ,群体倍增时间为 66h。Gomori萘酚磷酸酯法染色显示其胞浆内富含碱性磷酸酶颗粒 ,vonKossa染色表明聚集的细胞团能形成矿化结节。结论 :脂肪基质细胞中的成体干细胞经过矿化诱导培养后可向成骨细胞分化 ,并具有明显的成骨表型更多还原

【Abstract】 Objective: To isolate and culture human adipose tissu e derived stromal cells and to study their osteogenic potential. Meth ods: Human adipose tissue obtained by liposuction was minced and dige sted with collagenase typeⅠ. The obtained cells were plated in BGJ b medium i n primary culture for ten days. Following culture of the cells was in DMEM F12 medium supplemented with φ=10% fetal boven serum, 50 mg/L freshly prepared asc orbic acid,10 nmol/L vitamin D 3, 10 mmol/L Na β glycerophosphate and 10 - 8 mol/L dexamethasone for another 14 days as osteoblast inducing culture. The cells were observed under inverted microscope each day and cell growth was stud ied with cell counting. The osteoblast phenotype was assayed with Gomori and Von Kossa staining. Results: Polygon shaped cells were chara cterized by large size with a relatively large nucleus and plenty cytoplasm aft er osteo inducing cultivation. The population doubling time of the cells was 66 hours. Gomori and Von Kossa staining demonstrated positive expression of ALP an d formation of mineralized nods in extracellular matrix. Conclusion: It seems that human adipose tissue represents an abundant reservoir o f stem cells with the potential of osteoblast differentiation.更多还原