Munir Samir, Foldager Casper Bindzus, Lind Martin, Zachar Vladimir, Søballe Kjeld, Koch Thomas Gadegaard
Orthopaedic Research Laboratory, Institute for Clinical Medicine, Aarhus University Hospital, Noerrebrogade 44 Bldg 1a, 8000, Aarhus C, Denmark.
Cell Tissue Res. 2014 Jan;355(1):89-102. doi: 10.1007/s00441-013-1732-5. Epub 2013 Nov 1.
Human adipose-derived stromal cells (hASCs) possess the potential for chondrogenic differentiation. Recent studies imply that this differentiation process may be enhanced by culturing the cells in low oxygen tension in combination with three-dimensional (3D) scaffolds. We report the evaluation of the chondrogenic potential of hASC pellets in 5 and 21% O2 and as cell-scaffold constructs using a collagen I/III scaffold with chemical induction using TGF-β3. hASCs from four human donors were cultured both in a micromass pellet system and in 3D collagen I/III scaffolds in either 5 or 21% O2. Chondrogenesis was evaluated by quantitative gene expression analysis of aggrecan, SOX9, collagen I, II and X and histological evaluation with H&E and toluidine blue staining. Induced pellets cultured in 5% O2 showed increased peripheral cellularity and matrix deposition compared with 21% O2. Induced pellets cultured in 5% O2 had increased control-adjusted gene expression of aggrecan, SOX9 and collagen I and decreased collagen X compared with 21% O2 cultures. Induced pellets had higher gene expression of aggrecan, SOX9, collagen I, II and X and increased ratios of collagen II/I and collagen II/X compared with controls. As for pellets, scaffold cultures showed cellularity and matrix deposition organized in a zonal manner as a function of the oxygen tension, with a cartilage-like morphology and matrix deposition peripherally in the 5% O2 group and a more centrally located matrix in the 21% O2 group. There were no differences in histology and gene expressions between pellet and scaffold cultures. Five percent O2 in combination with chondrogenic culture medium stimulated chondrogenic differentiation of hASCs in vitro. We observed similar patterns of differentiation and matrix disposition in pellet and scaffold cultures.
人脂肪来源的基质细胞（hASC）具有软骨形成分化的潜力。最近的研究表明，通过在低氧张力下结合三维（3D）支架培养细胞，这种分化过程可能会增强。我们报告了在5%和21%氧气条件下对hASC微球软骨形成潜力的评估，以及使用I/III型胶原支架并通过TGF-β3化学诱导构建细胞-支架结构的评估。来自四名人类供体的hASC在微团粒系统以及5%或21%氧气条件下的3D I/III型胶原支架中培养。通过对聚集蛋白聚糖、SOX9、I、II和X型胶原进行定量基因表达分析以及用苏木精和伊红（H&E）染色和甲苯胺蓝染色进行组织学评估来评价软骨形成。与在21%氧气条件下培养相比，在5%氧气条件下培养的诱导微球显示外周细胞数量增加和基质沉积增加。与在21%氧气条件下培养的诱导微球相比，在5%氧气条件下培养的诱导微球经对照调整后的聚集蛋白聚糖、SOX9和I型胶原基因表达增加，而X型胶原基因表达降低。与对照相比，诱导微球的聚集蛋白聚糖、SOX9、I、II和X型胶原基因表达更高，且II/I型胶原和II/X型胶原比率增加。对于微球，支架培养显示细胞数量和基质沉积根据氧张力呈区域化组织，在5%氧气组中具有类似软骨的形态且基质沉积在外周，而在21%氧气组中基质更位于中央。微球培养和支架培养在组织学和基因表达方面没有差异。5%氧气与软骨形成培养基联合刺激了hASC在体外的软骨形成分化。我们在微球培养和支架培养中观察到了相似的分化模式和基质分布。
