Department of Veterinary Science, School of Veterinary medicine, Nippon Veterinary and Life Science University, 1-7-1 Kyonancho, Musashino, Tokyo 180-8602, Japan.
BMC Vet Res. 2012 Aug 31;8:150. doi: 10.1186/1746-6148-8-150.
Bone marrow-derived mesenchymal stem cells (BM-MSCs) and adipose tissue-derived mesenchymal stem cells (AT-MSCs) are potential cellular sources of therapeutic stem cells. MSCs are a multipotent population of cells capable of differentiating into a number of mesodermal lineages. Treatment using MSCs appears to be a helpful approach for structural restoration in regenerative medicine. Correct identification of these cells is necessary, but there is inadequate information on the MSC profile of cell surface markers and mRNA expression in dogs. In this study, we performed molecular characterization of canine BM-MSCs and AT-MSCs using immunological and mRNA expression analysis.
Samples were confirmed to be multipotent based on their osteogenic and adipogenic differentiation. And these cells were checked as stem cell, hematopoietic and embryonic stem cell (ESC) markers by flow cytometry. BM- and AT-MSCs showed high expression of CD29 and CD44, moderate expression of CD90, and were negative for CD34, CD45, SSEA-3, SSEA-4, TRA-1-60, and TRA-1-81. SSEA-1 was expressed at very low levels in AT-MSCs. Quantitative real-time PCR (qRT-PCR) revealed expression of Oct3/4, Sox2, and Nanog in BM- and AT-MSCs. There was no significant difference in expression of Oct3/4 and Sox2 between BM-MSCs and AT-MSCs. However, Nanog expression was 2.5-fold higher in AT-MSCs than in BM-MSCs. Using immunocytochemical analysis, Oct3/4 and Sox2 proteins were observed in BM- and AT-MSCs.
Our results provide fundamental information to enable for more reproducible and reliable quality control in the identification of canine BM-MSCs and AT-MSCs by protein and mRNA expression analysis.
骨髓间充质干细胞（BM-MSCs）和脂肪组织来源的间充质干细胞（AT-MSCs）是治疗性干细胞的潜在细胞来源。MSCs 是一种多能细胞群，能够分化为许多中胚层谱系。使用 MSCs 进行治疗似乎是再生医学中结构修复的一种有益方法。正确识别这些细胞是必要的，但目前关于犬 MSC 表面标志物和 mRNA 表达的信息不足。在这项研究中，我们使用免疫和 mRNA 表达分析对犬 BM-MSCs 和 AT-MSCs 进行了分子特征分析。
基于成骨和成脂分化，样本被确认为多能性。并且这些细胞通过流式细胞术被检查为干细胞、造血细胞和胚胎干细胞（ESC）标志物。BM-MSCs 和 AT-MSCs 均高表达 CD29 和 CD44，中度表达 CD90，且阴性表达 CD34、CD45、SSEA-3、SSEA-4、TRA-1-60 和 TRA-1-81。SSEA-1 在 AT-MSCs 中的表达水平非常低。实时定量 PCR（qRT-PCR）显示 BM-MSCs 和 AT-MSCs 中表达 Oct3/4、Sox2 和 Nanog。BM-MSCs 和 AT-MSCs 之间 Oct3/4 和 Sox2 的表达没有显著差异。然而，Nanog 的表达在 AT-MSCs 中是 BM-MSCs 的 2.5 倍。通过免疫细胞化学分析，在 BM-MSCs 和 AT-MSCs 中观察到 Oct3/4 和 Sox2 蛋白。
我们的结果提供了基本信息，通过蛋白质和 mRNA 表达分析，可更准确地识别犬 BM-MSCs 和 AT-MSCs，从而实现更可重复和可靠的质量控制。
