【作者】 林云锋；

【导师】 田卫东；

【作者基本信息】 四川大学 ， 口腔临床医学， 2003， 硕士

【摘要】 脂肪基质中存在一些具有多向分化潜能的具有自我更新和一定分化潜能，处于全能干细胞和成熟脂肪细胞之间的成体干细胞，具有向脂肪细胞、成骨细胞、成软骨细胞、肌细胞和神经细胞等多个方向分化的能力。 本实验由三部分组成： （1）脂肪基质细胞的获取及培养 采用抽脂术获得的健康成体脂肪组织，经机械分割、胶原酶消化后，BGJ_b培养基培养、传代。 （2）脂肪基质细胞成骨诱导培养 以DMEM／F12培养基提供脂肪基质细胞向成骨方向分化所需要的生长因子；β—磷酸甘油钠提供形成矿化基质提供所需的磷酸根离子和钙离子；地塞米松抑制细胞的增殖而加速细胞的分化；左旋抗坏血酸及VitD₃可以诱导脂肪基质细胞向成骨方向分化。 （3）以光镜观察、常规HE染色、Von Kossa’s矿化结节染色法、Gomori改良钙钴法ALP染色、Gomori萘酚磷酸酯法等方法对所获得之矿化诱导后的脂肪间质细胞进行鉴定。 研究结果表明：人体脂肪基质细胞在诱导条件下，可以分化成为成骨细胞样细胞，细胞周围可以形成类似于羟基磷灰石样的磷酸钙盐的沉积物，并可以形成矿化结节样的结构。而且可以表现出较高碱性磷酸酶（ALP）活性。 实验证明，在人类脂肪组织的基质细胞中，存在具有成骨方向分化潜能的成体干细胞；在矿化诱导条件下，人类成体脂肪组织的基质细胞可以向成骨方向转化，可以形成具有成骨表型的细胞。这些研究可以为下一步人成体脂肪基质细胞在骨组织工程的应用提供实验基础。更多还原

【Abstract】 It is hypothesized that human adipose tissue is an abundant reservoir of adult stem cells, which have potentent to differentiate into many mature cell lineages varying from adipogenic precursors, osteogenic precursors, chondrogenic precursors to myogenic precursors.This research includes three main parts:(1) Isolation and cultivation: Human subcutaneous adipose tissue was acquired by liposuction. The tissue was then digested with one volume of collagenase type I and cultured with BGJb medium.(2) Mineralizing induction: Human adipose tissue-derived stromal cells were cultured and differentiated into osteoblasts in midium of DMEM/F12, supplemented with L-ascorbate, β-glycerophosphate, dexamethasone, 1,25-dihydroxy vitamin D3.(3) Histochemical stains: HE staining, Von Kossa’s histochemical staining and alkaline phosphatase assay were employed to identify the osteogenic phenotypes of human adipose tissue-derived stromal cells after osteogenic induction.The results of this research showed: Human adipose tissue -derived stromal cells were able to differentiate into osteoblasts under condition of mineralizing induction. The alkaline phosphatase activity and extracellular calcified nodes were positive among these cells.It seems that human adipose tissue is of adult stem cells, which with potential to differentiate into osteoblasts in vitro. This study may provide some practical methods for engineering bone in future.更多还原