A novel serum-free medium for the expansion of human mesenchymal stem cells.
A novel serum-free medium for the expansion of human mesenchymal stem cells.
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DOI：
10.1186/scrt8
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发表时间：
2010-04-02
影响因子：
7.5
通讯作者：
Vemuri MC
中科院分区：
文献类型：
--
作者：
Chase LG;Lakshmipathy U;Solchaga LA;Rao MS;Vemuri MC
人类多能间充质干细胞（MSC）疗法正在临床上测试各种疾病，包括克罗恩病，多发性硬化症，移植物抗宿主病，1型糖尿病，骨折和软骨缺损。然而，尽管在该领域取得了显著的临床进展，但大多数应用仍然使用含有胎牛血清的传统培养基。传统培养基的不明确和高度可变的性质仍然是一个挑战，阻碍了基础和临床人类MSC研究领域。迄今为止，还没有可靠的用于人MSC的无血清培养基。在这项研究中，我们开发并测试了无血清生长培养基对人骨髓来源的间充质干细胞通过调查的多个参数，包括原代细胞分离，多次传代扩增，中胚层分化，细胞表型，基因表达分析。与传统培养基相似，在补充有重组人血小板衍生生长因子-BB（PDGF-BB）、碱性成纤维细胞生长因子（bFGF）和转化生长因子（TGF）-β1的无血清培养基中扩增的人MSC显示出广泛的增殖，保留了表型、分化和集落形成单位潜力。为了监测全局基因表达，比较了在无血清和含血清条件下扩增的骨髓来源的MSC的转录组，揭示了相似的表达谱。此外，所述的无血清培养基支持从原代人骨髓抽吸物中分离人MSC并持续增殖。尽管所述的无血清MSC培养基并非不含异种成分，但该培养基为研究应用提供了含血清培养基的替代品，为未来的临床应用奠定了基础。
Human multipotent mesenchymal stem cell (MSC) therapies are being tested clinically for a variety of disorders, including Crohn's disease, multiple sclerosis, graft-versus-host disease, type 1 diabetes, bone fractures, and cartilage defects. However, despite the remarkable clinical advancements in this field, most applications still use traditional culture media containing fetal bovine serum. The ill-defined and highly variable nature of traditional culture media remains a challenge, hampering both the basic and clinical human MSC research fields. To date, no reliable serum-free medium for human MSCs has been available. In this study, we developed and tested a serum-free growth medium on human bone marrow-derived MSCs through the investigation of multiple parameters including primary cell isolation, multipassage expansion, mesoderm differentiation, cellular phenotype, and gene-expression analysis. Similar to that achieved with traditional culture medium, human MSCs expanded in serum-free medium supplemented with recombinant human platelet-derived growth factor-BB (PDGF-BB), basic fibroblast growth factor (bFGF), and transforming growth factor (TGF)-β1 showed extensive propagation with retained phenotypic, differentiation, and colony-forming unit potential. To monitor global gene expression, the transcriptomes of bone marrow-derived MSCs expanded under serum-free and serum-containing conditions were compared, revealing similar expression profiles. In addition, the described serum-free culture medium supported the isolation of human MSCs from primary human marrow aspirate with continual propagation. Although the described serum-free MSC culture medium is not free of xenogeneic components, this medium provides a substitute for serum-containing medium for research applications, setting the stage for future clinical applications.
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