This study was to determine the ectopic osteogenic ability of BMSCs in combination with a scaffolding material comprising hydroxyapatite and β-tricalcium phosphate matrix (HA/β-TCP). BMSCs were obtained from the SD rats and induced to osteogenesis. Then these induced cells were seeded into HA/β-TCP and the constructs were auto-implanted subcutaneously for up to 12 weeks. Histological analysis, immunostaing, RT-PCR and transmission electron microscopy of the retrieved specimens at various intervals showed obvious trends of ectopic bone formation with obvious alteration of cellular phenotype.

Musculoskeletal tissues regeneration requires rapid expansion of seeding cells both in vitro and in vivo while maintaining their multilineage differentiation ability. Human adipose-derived stem cells (ASCs) are considered to contain multipotent mesenchymal stem cells. Monolayer cultures of human ASCs were isolated from human lipoaspirates and passaged 3 times and then infected with replication-incompetent adenoviral vectors carrying green fluorescent protein (Ad/GFP) genes. Then, Ad/GFP infected human ASCs were transferred to osteogenic, chondrogenic, adipogenic, and myogenic medium. The morphological characterization of induced cells was observed using phase-contrast microscopy and fluorescence microscopy. The expression of marker proteins or genes was measured by immunocytochemical and RT-PCR analysis. Osteopontin (OPN), and osteocalcin (OCN) were positive in osteogenic lineages, aggrecan and SOX9 were positive in chondrogenic ones, peroxisome proliferatoractivated receptor (PPAR-γ 2) and lipoprotein lipase (LPL) were positive in adipogenic ones, and myogenin and myod1 was positive in myogenic ones. At the same time, the results of fluorescence microscopic imaging proved that the high level of GFP expression during ASCs differentiation maintained stable nearly 2 months. So the exogenous GFP and multilineage potential of human ASCs had no severe influences on each other. Since the human ASCs can be easily obtained and abundant, it is proposed that they may be promising candidate cells for further studies on tissue engineering. Imaging with expression of GFP facilitates the research on ASCs physiological behavior and application in tissue engineering during differentiation both in vitro and in vivo.

分离培羊人体脂肪基质细胞，经过软骨定向诱导后复合藻酸盐凝胶，研究基异位软骨生成能力。方法将脂肪抽吸术获取的人体脂肪机械分割，通过I型胶原酶消化后得到脂肪10ug/LTCF-β，50mg/L维生素C的DMEM/F12培羊基中诱导培羊14d，获取的细胞按1×1010/L的细胞密度与质量分数为1.2%的藻酸钠复合、加入102mmol/LCaCl，充分泥匀，使之交联，将细胞-藻酸盐凝胶1ml注入每只体重为25g的BALB/C裸小隶鼠背部皮下（共4办），同时在裸小鼠臀部两侧皮下各注入相同量的无细胞藻酸盐及单纯细胞作为自岙对照、术后4、8周各处列2只动物，取材固定、脱钙、包埋后切自染色。镜下观察。结果经连续导培羊14d的脂肪基质细胞已具明显成骨表型（细胞基质中富含硫酸软骨素及II型胶原）。4、8周时，注射细胞-藻酸盐胶标本均显示成明显的软骨形成。肥大的软骨细非金属位于富含硫在术手3-4周已完全吸收。结论脂肪基质细胞过导培羊具有向软骨细胞表型分化的潜能。

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/Lβ2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。

目的研究分离的人体脂肪基质细胞经过诱导培养后向骨骼肌细胞分化的潜能。方法通过脂肪抽吸术获取健康成人脂肪，机械分割后用Ⅰ型胶原酶消化，得到脂肪基质细胞。将脂肪基质细胞在BGJb培养基中原代培养10d，在DMEM/F12培养基中进行成肌诱导培养20d，获得细胞爬片。细胞爬片经4%多聚甲醛固定后，用甲苯胺蓝、Mallory磷钨酸苏木素染色观察细胞形态；Myosin单克隆抗体免疫细胞化学染色观察肌球蛋白表达情况。结果经过成肌诱导培养的细胞与常规培养的脂肪基质细胞在形态上有明显的差异，表现为细胞体积增大，单个核的细胞融合形成多核的肌管样结构，胞浆内细胞器发达，肌浆网扩张并形成肌原纤维骨骼肌特异性的Myosin表达阳性。未经诱导的脂肪基质细胞在相同时间点无此现象。结论人类脂肪基质细胞中存在着成体干细胞，在成肌诱导培养条件下具有向骨骼肌细胞分化的潜能。