Characterization and multipotentiality of human fetal femur-derived cells: Implications for skeletal tissue regeneration
Characterization and multipotentiality of human fetal femur-derived cells: Implications for skeletal tissue regeneration
复制标题
DOI：
10.1634/stemcells.2005-0368
复制
发表时间：
2006-04-01
期刊：
影响因子：
5.2
通讯作者：
Oreffo, Richard O. C.
中科院分区：
文献类型：
--
作者：
Mirmalek-Sani, Sayed-Hadi;Tare, Rahul S.;Oreffo, Richard O. C.
迄今为止，胚胎骨骼组织祖细胞的可塑性、多能性和特征仍不清楚。本研究对人胎儿股骨的细胞群与成人间充质细胞群进行了比较。实时定量聚合酶链反应显示，与成人来源的和免疫选择的STRO 1富集的成人群体相比，胎儿来源的细胞表达间充质祖细胞标志物。使用来自股骨和单细胞克隆的细胞检查多潜能性，从外植体培养扩增，并在暴露于成脂、成骨和成软骨条件之前维持在基础培养基中。通过油红0脂质染色和aP 2免疫细胞化学证实了脂肪细胞的形成，仅在成脂条件下检测到过氧化物酶体增殖激活受体γ的表达。在成软骨细胞团中，用阿尔新蓝/天狼星红染色和II型胶原免疫细胞化学观察到软骨细胞嵌入陷窝和包埋在致密的蛋白多糖基质中。成骨分化通过碱性磷酸酶染色和I型胶原免疫细胞化学以及骨桥蛋白和骨钙素的基因表达来证实。使用单细胞克隆分析来证明胎儿来源的群体的多潜能性，形成脂肪形成、软骨形成和成骨群体。在严重受损的免疫缺陷小鼠皮下植入后，在体外和体内均观察到在仿生支架上培养后的矿化和类骨质形成，具有广泛的基质积累。这些研究表明，与成人来源的细胞相比，胎儿股骨来源的细胞的增殖和多潜能特性。选择性分化和免疫分型将决定这些胎儿细胞作为修复受损组织的独特替代模型和细胞来源的潜力。
To date, the plasticity, multipotentiality, and characteristics of progenitor cells from fetal skeletal tissue remain poorly defined. This study has examined cell populations from human fetal femurs in comparison with adult-derived mesenchymal cell populations. Real-time quantitative polymerase chain reaction demonstrated expression of mesenchymal progenitor cell markers by fetal-derived cells in comparison with unselected adult-derived and immunoselected STRO1-enriched adult populations. Multipotentiality was examined using cells derived from femurs and single-cell clones, culture-expanded from explants, and maintained in basal medium prior to exposure to adipogenic, osteogenic, and chondrogenic conditions. Adipocyte formation was confirmed by Oil Red 0 lipid staining and aP2 immunocytochemistry, with expression of peroxisome proliferation-activated receptor-gamma detected only in adipogenic conditions. In chondrogenic pellets, chondrocytes lodged within lacunae and embedded within dense proteoglycan matrix were observed using Alcian blue/Sirius red staining and type II collagen inuntmocytochemistry. Osteogenic differentiation was confirmed by alkaline phosphatase staining and type I collagen immunocytochemistry as well as by gene expression of osteopontin and osteocalcin. Single-cell clonal analysis was used to demonstrate multipotentiality of the fetal-derived populations with the formation of adipogenic, chondrogenic, and osteogenic populations. Mineralization and osteoid formation were observed after culture on biomimetic scaffolds with extensive matrix accumulation both in vitro and in vivo after subcutaneous implantation in severely compromised immunodeficient mice. These studies demonstrate the proliferative and multipotential properties of fetal femur-derived cells in comparison with adult-derived cells. Selective differentiation and immunophenotyping will determine the potential of these fetal cells as a unique alternative model and cell source in the restoration of damaged tissue.