Wu Ling, Prins Henk-Jan, Leijten Jeroen, Helder Marco N, Evseenko Denis, Moroni Lorenzo, van Blitterswijk Clemens A, Lin Yunfeng, Karperien Marcel
1 Department of Developmental BioEngineering, MIRA-Institute for Biomedical Technology and Technical Medicine, University of Twente , Enschede, The Netherlands .
2 Department of Orthopedic Surgery, Orthopedic Hospital Research Center, David Geffen School of Medicine, University of California , Los Angeles, California.
Tissue Eng Part A. 2016 Feb;22(3-4):336-48. doi: 10.1089/ten.TEA.2015.0269. Epub 2016 Feb 1.
Partial replacement of chondrocytes by stem cells has been proposed to improve the performance of autologous chondrocyte implantation (ACI). Our previous studies showed that the increased cartilage production in pellet cocultures of chondrocytes and mesenchymal stem cells (MSCs) is due to a trophic role of the MSCs by stimulating chondrocyte proliferation and matrix production rather than MSCs actively undergoing chondrogenic differentiation. The aim of this study is to compare the trophic effects of stromal vascular fraction cells (SVF) and in vitro expanded adipose stem cells (ASC). SVF and culture-expanded ASC (n = 9) were cocultured with primary human chondrocytes in pellets. By glycosaminoglycan (GAG) and DNA assays, we showed that coculture pellets of SVF and chondrocytes have more GAG deposition than that of ASC and chondrocytes. Results of the short tandem repeats analysis indicated that the increase in the chondrocyte proportion in the coculture pellets is more pronounced in the SVF coculture group than in the ASC coculture group. Using flow cytometry and microarray, we demonstrated that SVF and ASC have different characteristics in cell surface markers and gene expression profiles. SVF is more heterogeneous than ASC, whereas ASC is more enriched in cells from the mesenchymal lineage than SVF. By subcutaneous implantation into nude mice, we showed that constructs of SVF and chondrocytes are better in depositing cartilage matrix than the mixture of ASC and chondrocytes. Taken together, SVF is better than ASC in terms of forming cartilage matrix in pellet coculture and in coimplantation models omitting the need for prior cell expansion. Our study suggests that the SVF in combination with primary human chondrocytes may be a good cell combination for one-stage cartilage repair.
有人提出用干细胞部分替代软骨细胞以提高自体软骨细胞移植(ACI)的效果。我们之前的研究表明,软骨细胞与间充质干细胞(MSC)的微团共培养中软骨生成增加是由于MSC通过刺激软骨细胞增殖和基质产生发挥了营养作用,而非MSC积极地进行软骨分化。本研究的目的是比较基质血管成分细胞(SVF)和体外扩增的脂肪干细胞(ASC)的营养作用。将SVF和培养扩增的ASC(n = 9)与原代人软骨细胞进行微团共培养。通过糖胺聚糖(GAG)和DNA检测,我们发现SVF与软骨细胞的共培养微团比ASC与软骨细胞的共培养微团有更多的GAG沉积。短串联重复序列分析结果表明,共培养微团中软骨细胞比例的增加在SVF共培养组比ASC共培养组更明显。使用流式细胞术和微阵列,我们证明SVF和ASC在细胞表面标志物和基因表达谱方面具有不同特征。SVF比ASC更具异质性,而ASC比SVF在间充质谱系细胞中更富集。通过皮下植入裸鼠,我们发现SVF与软骨细胞构建体在沉积软骨基质方面比ASC与软骨细胞的混合物更好。综上所述,在微团共培养以及无需预先细胞扩增的共植入模型中,就形成软骨基质而言,SVF优于ASC。我们的研究表明,SVF与原代人软骨细胞联合可能是一期软骨修复的良好细胞组合。