Teunissen M, Verseijden F, Riemers F M, van Osch G J V M, Tryfonidou M A
Department of Clinical Sciences, Faculty of Veterinary Medicine, Utrecht University, Yalelaan 108, 3584 CM, Utrecht, The Netherlands.
Department of Clinical Sciences, Faculty of Veterinary Medicine, Utrecht University, Yalelaan 108, 3584 CM, Utrecht, The Netherlands.
Vet J. 2021 Mar;269:105605. doi: 10.1016/j.tvjl.2020.105605. Epub 2020 Dec 29.
Mesenchymal stromal cells (MSC) are used for cell-based treatment for canine osteoarthritis (OA). Compared with human MSCs, detailed information on the functional characterisation of canine MSCs is limited. In particular, the chondrogenic differentiation of canine adipose tissue-derived MSCs (cAT-MSCs) is challenging. In this study, we aimed to compare cAT-MSCs with bone marrow-derived MSCs (cBM-MSCs), focusing specifically on their in vitro chondrogenic potential, with or without bone morphogenetic proteins (BMP). cBM-MSCs and cAT-MSCs were characterised using flow cytometry and reverse transcription-quantitative polymerase chain reaction (RT-qPCR). The chondrogenic differentiation potential of all cMSC preparations in the presence of TGF-β1 alone or when supplemented with 10, 100, or 250 ng/mL BMP-2 or BMP-6 was investigated using RT-qPCR, and biochemical, histochemical and immunohistological analyses. Both cBM-MSCs and cAT-MSCs expressed the surface markers CD90, CD73, and CD29, and were negative for CD45 and CD34, although the expression of CD73 and CD271 varied with donor and tissue origin. Interestingly, expression of ACAN and SOX9 was higher in cBM-MSCs than cAT-MSCs. In contrast with cBM-MSCs, cAT-MSCs could not differentiate toward the chondrogenic lineage without BMP-2/-6, and their in vitro chondrogenesis was inferior to cBM-MSCs with BMP-2/-6. Thus, cAT-MSCs have lower in vitro chondrogenic capacity than cBM-MSC under the studied culture conditions with 10, 100, or 250 ng/mL BMP-2 or BMP-6. Therefore, further characterisation is necessary to explore the potential of cAT-MSCs for cell-based OA treatments.
间充质基质细胞（MSC）被用于犬骨关节炎（OA）的细胞治疗。与人类MSC相比，关于犬MSC功能特性的详细信息有限。特别是，犬脂肪组织来源的MSC（cAT-MSC）的软骨形成分化具有挑战性。在本研究中，我们旨在比较cAT-MSC与骨髓来源的MSC（cBM-MSC），特别关注它们在有或没有骨形态发生蛋白（BMP）情况下的体外软骨形成潜力。使用流式细胞术和逆转录定量聚合酶链反应（RT-qPCR）对cBM-MSC和cAT-MSC进行表征。使用RT-qPCR以及生化、组织化学和免疫组织学分析，研究了所有cMSC制剂在单独存在转化生长因子-β1（TGF-β1）或补充10、100或250 ng/mL BMP-2或BMP-6时的软骨形成分化潜力。cBM-MSC和cAT-MSC均表达表面标志物CD90、CD73和CD29，而CD45和CD34呈阴性，尽管CD73和CD271的表达因供体和组织来源而异。有趣的是，cBM-MSC中ACAN和SOX9的表达高于cAT-MSC。与cBM-MSC不同，cAT-MSC在没有BMP-2/-6的情况下不能向软骨形成谱系分化，并且它们的体外软骨形成能力低于添加BMP-2/-6的cBM-MSC。因此，在研究的添加10、100或250 ng/mL BMP-2或BMP-6的培养条件下，cAT-MSC的体外软骨形成能力低于cBM-MSC。因此，有必要进一步表征以探索cAT-MSC在基于细胞的OA治疗中的潜力。
