Stem Cell Research Laboratory, Section of Hematology, Department of Medicine, Policlinico G.B. Rossi-University of Verona, Verona, Italy.
Stem Cells Dev. 2010 Oct;19(10):1449-70. doi: 10.1089/scd.2010.0140.
Mesenchymal stem cells (MSCs) are adult stem cells that hold great promise in the field of regenerative medicine. They can be isolated from almost any tissue of the body and display, after expansion, very similar properties and minor differences, probably due to their microenvironment of origin. Expansion in vitro can be obtained in cytokine-free, serum-enriched media, as well as in serum-free, basic fibroblast growth factor-enriched media. A detailed immunophenotypic analysis is required to test the purity of the preparation, but no unique distinguishing marker has been described as yet. Functional assays, that is, differentiation studies in vitro, are needed to prove multilineage differentiation of expanded cells, and demonstration of pluripotency is necessary to identify most immature precursors. MSCs show powerful immunomodulative properties toward most of the cells of the immune system: this strengthens the theoretical rationale for their use also in an allogeneic setting across the major histocompatibility complex (MHC) immunological barriers. Systemic intravenous injection and local use have been tried: after systemic injection, MSCs show a high degree of chemotaxis based on pro-inflammatory cytokines, and localize at inflamed and neoplastic tissues; local regeneration has been improved using synthetic, as well as organic scaffolds. On the other hand, inadequate heterotopic in vivo differentiation and neoplastic transformation are potential risks of this form of cell therapy, even if evidence of this sort has been collected only from studies in mice, and generally after prolonged in vitro expansion. This review tries to provide a detailed technical overview of the methods used for human bone-marrow (BM)-derived and adipose-tissue (AT)-derived MSC isolation, in vitro expansion, and characterization for tissue repair. We chose to use BM-MSCs as a model to describe techniques that have been used for MSC isolation and expansion from very different sources, and AT-MSCs as an example of a reliable and increasingly common alternative source.
间充质干细胞（MSCs）是成体干细胞，在再生医学领域具有巨大的应用前景。它们可以从身体的几乎任何组织中分离出来，经过扩增后具有非常相似的特性和微小的差异，这可能是由于它们起源的微环境所致。在无细胞因子、富含血清的培养基以及无血清、富含碱性成纤维细胞生长因子的培养基中均可进行体外扩增。需要进行详细的免疫表型分析来测试制剂的纯度，但尚未描述任何独特的鉴别标志物。需要进行功能测定，即体外分化研究，以证明扩增细胞的多能性分化，并证明多能性分化以鉴定大多数未成熟的前体细胞。MSCs 对免疫系统的大多数细胞都具有强大的免疫调节特性：这加强了其在同种异体背景下跨越主要组织相容性复合物（MHC）免疫屏障的理论依据。已尝试进行全身静脉内注射和局部应用：全身注射后，MSCs 根据促炎细胞因子表现出高度趋化性，并定位于炎症和肿瘤组织；使用合成和有机支架改善了局部再生。另一方面，这种细胞疗法存在异位体内分化不足和肿瘤转化的潜在风险，尽管这种风险的证据仅来自于小鼠研究，并且通常是在长时间的体外扩增之后才出现。本综述试图提供关于人骨髓（BM）衍生和脂肪组织（AT）衍生 MSC 分离、体外扩增和组织修复特性的详细技术概述。我们选择使用 BM-MSCs 作为模型来描述从非常不同来源分离和扩增 MSC 所使用的技术，并以 AT-MSCs 为例说明一种可靠且越来越常见的替代来源。
