Development and Characterization of a Clinically Compliant Xeno-Free Culture Medium in Good Manufacturing Practice for Human Multipotent Mesenchymal Stem Cells
Development and Characterization of a Clinically Compliant Xeno-Free Culture Medium in Good Manufacturing Practice for Human Multipotent Mesenchymal Stem Cells
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DOI：
10.5966/sctm.2012-0072
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发表时间：
2012-10-01
影响因子：
6
通讯作者：
Vemuri, Mohan C.
中科院分区：
文献类型：
--
作者：
Chase, Lucas G.;Yang, Sufang;Vemuri, Mohan C.
人类多能间充质干细胞（MSC）疗法目前正在克罗恩病，多发性硬化症，移植物抗宿主病，1型糖尿病，骨折，软骨损伤和心脏病的临床试验中进行测试。尽管在临床试验中取得了显著进展，但大多数应用仍然使用含有胎牛血清的传统培养基或含有血清白蛋白、胰岛素和转铁蛋白的无血清培养基。传统培养基的不明确和可变性质仍然是一个挑战，并且需要更好地定义无异种培养基，以满足基于细胞的疗法的监管和长期安全性要求。我们开发并测试了一种无血清和无异种的培养基（SFM-XF），使用人骨髓和脂肪来源的MSC，通过研究原代细胞分离，多次传代扩增，中胚层分化，细胞表型和基因表达分析，这对于遵守翻译到细胞治疗至关重要。在SFM-XF中扩增的人MSC显示出持续的增殖，具有预期的表型和向成脂、成软骨和成骨谱系的分化潜力，与在传统含血清培养基（SCM）中扩增的MSC相似。为了监测整体基因表达，比较了在SFM-XF和SCM中扩增的骨髓源性MSC的转录组，揭示了相对相似的表达谱。此外，SFM-XF支持从原代人骨髓抽吸物中分离和增殖人MSC，确保这些方法和试剂适用于转化为治疗。SFM-XF培养系统允许MSC的更好的扩增和多潜能性，并作为含血清培养基的优选替代品，用于生产大规模的、功能上胜任的MSC以用于未来的临床应用。干细胞转化医学2012;1：750-758
Human multipotent mesenchymal stem cell (MSC) therapies are currently being tested in clinical trials for Crohn's disease, multiple sclerosis, graft-versus-host disease, type 1 diabetes, bone fractures, cartilage damage, and cardiac diseases. Despite remarkable progress in clinical trials, most applications still use traditional culture media containing fetal bovine serum or serum-free media that contain serum albumin, insulin, and transferrin. The ill-defined and variable nature of traditional culture media remains a challenge and has created a need for better defined xeno-free culture media to meet the regulatory and long-term safety requirements for cell-based therapies. We developed and tested a serum-free and xeno-free culture medium (SFM-XF) using human bone marrow- and adipose-derived MSCs by investigating primary cell isolation, multiple passage expansion, mesoderm differentiation, cellular phenotype, and gene expression analysis, which are critical for complying with translation to cell therapy. Human MSCs expanded in SFM-XF showed continual propagation, with an expected phenotype and differentiation potential to adipogenic, chondrogenic, and osteogenic lineages similar to that of MSCs expanded in traditional serum-containing culture medium (SCM). To monitor global gene expression, the transcriptomes of bone marrow-derived MSCs expanded in SFM-XF and SCM were compared, revealing relatively similar expression profiles. In addition, the SFM-XF supported the isolation and propagation of human MSCs from primary human marrow aspirates, ensuring that these methods and reagents are compatible for translation to therapy. The SFM-XF culture system allows better expansion and multipotentiality of MSCs and serves as a preferred alternative to serum-containing media for the production of large scale, functionally competent MSCs for future clinical applications. STEM CELLS TRANSLATIONAL MEDICINE 2012;1:750-758