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髌下脂肪垫和皮下脂肪组织来源的与供体匹配的间充质干细胞共培养对人骨关节炎软骨细胞软骨生成的影响。

Influence on chondrogenesis of human osteoarthritic chondrocytes in co-culture with donor-matched mesenchymal stem cells from infrapatellar fat pad and subcutaneous adipose tissue.

机构信息

Cell and Tissue Engineering Laboratory, Gruppo Ospedaliero San Donato Foundation, Milan, Italy.

出版信息

Int J Immunopathol Pharmacol. 2013 Jan-Mar;26(1 Suppl):23-31. doi: 10.1177/03946320130260S104.

DOI:10.1177/03946320130260S104
PMID:24046946
Abstract

Co-culture of mesenchymal stem cells (MSCs) and articular chondrocytes (ACs) has been proposed for autologous cartilage cell-based therapies, to overcome the issues associated to limited availability of articular chondrocytes (ACs). To evaluate the potentiality of a co-culture approach in aged osteoarthritic patients, MSCs from infrapatellar fat pad (IFP-MSCs) and knee subcutaneous adipose tissue (ASCs) were co-cultured with donor-matched osteoarthritic, expanded and cryopreserved, ACs in a 75%/25% ratio. Co-cultures were prepared also from nasal chondrocytes (NCs) to evaluate their possible use as an alternative to ACs. Pellets were differentiated for 14 days, using mono-cultures of each cell type as reference. Chondrogenic genes SOX9, COL2A1, ACAN were less expressed in co-cultures compared to ACs and NCs. Total GAGs content in co-cultures did not differ significantly from values predicted as the sum of each cell type contribution corrected for the co-culture ratio, as confirmed by histology. No significant differences were observed for GAGs/DNA in mono-cultures, demonstrating a reduced chondrogenic potential of ACs and NCs. In conclusion, a small percentage of expanded and cryopreserved ACs and NCs did not lead to IFP-MSCs and ASCs chondro-induction. Our results suggest that chondrogenic potential and origin of chondrocytes may play a relevant role in the outcome of co-cultures, indicating a need for further investigations to demonstrate their clinical relevance in the treatment of aged osteoarthritic patients.

摘要

间充质干细胞 (MSCs) 和关节软骨细胞 (ACs) 的共培养已被提议用于自体软骨细胞为基础的治疗,以克服与关节软骨细胞 (ACs) 有限可用性相关的问题。为了评估共培养方法在老年骨关节炎患者中的潜力,来自髌下脂肪垫的间充质干细胞 (IFP-MSCs) 和膝关节皮下脂肪组织 (ASCs) 与供体匹配的、扩增和冷冻保存的 OA 软骨细胞以 75%/25% 的比例进行共培养。还从鼻软骨细胞 (NCs) 制备共培养物,以评估它们作为 ACs 的替代物的可能用途。使用每种细胞类型的单核培养物作为参考,对共培养物进行了 14 天的分化。与 ACs 和 NCs 相比,共培养物中软骨形成基因 SOX9、COL2A1 和 ACAN 的表达较低。共培养物中的总 GAGs 含量与根据共培养比例校正的每个细胞类型贡献的总和预测的值没有显著差异,这通过组织学得到证实。单核培养物中 GAGs/DNA 没有观察到显著差异,表明 ACs 和 NCs 的软骨形成潜力降低。总之,扩增和冷冻保存的 ACs 和 NCs 的一小部分并未导致 IFP-MSCs 和 ASCs 软骨诱导。我们的结果表明,软骨细胞的软骨形成潜力和来源可能在共培养物的结果中发挥重要作用,这表明需要进一步研究以证明它们在治疗老年骨关节炎患者中的临床相关性。

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