Three-dimensional spheroid culture of human umbilical cord mesenchymal stem cells promotes cell yield and stemness maintenance

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• 作者：Yi Li ; Gang Guo ; Li Li ; Fei Chen ; Ji Bao ; Yu-jun Shi ; Hong Bu
• 关键词：Mesenchymal stem cells ; hUC ; MSCs ; 3D culture ; Yield ; Stemness ; Human ; Mouse
• 刊名：Cell and Tissue Research
• 出版年：2015
• 出版时间：May 2015
• 年：2015
• 卷：360
• 期：2
• 页码：297-307
• 全文大小：6,302 KB
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• 作者单位：Yi Li (1) (2)
  Gang Guo (1) (2)
  Li Li (1)
  Fei Chen (1)
  Ji Bao (1) (2)
  Yu-jun Shi (1) (2)
  Hong Bu (2) (3)
  
  1. Laboratory of Pathology, West China Hospital, Sichuan University, Chengdu, 610041, China
  2. Key Laboratory of Transplant Engineering and Immunology, Ministry of Health, West China Hospital, Sichuan University, Chengdu, 610041, China
  3. Department of Pathology, Western China Hospital, Sichuan University, Chengdu, 610041, China
  
• 刊物类别：Biomedical and Life Sciences
• 刊物主题：Biomedicine
  Human Genetics
  Proteomics
  Molecular Medicine
  
• 出版者：Springer Berlin / Heidelberg
• ISSN：1432-0878

文摘

Mesenchymal stem cell (MSC) transplantation is a promising treatment of many diseases. However, conventional techniques with cells being cultured as a monolayer result in slow cell proliferation and insufficient yield to meet clinical demands. Three-dimensional (3D) culture systems are gaining attention with regard to recreating a complex microenvironment and to understanding the conditions experienced by cells. Our aim is to establish a novel 3D system for the culture of human umbilical cord MSCs (hUC-MSCs) within a real 3D microenvironment but with no digestion or passaging. Primary hUC-MSCs were isolated and grown in serum-free medium (SFM) on a suspension Rocker system. Cell characteristics including proliferation, phenotype and multipotency were recorded. The therapeutic effects of 3D-cultured hUC-MSCs on carbon tetrachloride (CCl₄)-induced acute liver failure in mouse models were examined. In the 3D Rocker system, hUC-MSCs formed spheroids in SFM and maintained high viability and active proliferation. Compared with monolayer culture, the 3D-culture system yielded more hUC-MSCs cells within the same volume. The spheroids expressed higher levels of stem cell markers and displayed stronger multipotency. After transplantation into mouse, 3D hUC-MSCs significantly promoted the secretion of interferon-γ and interleukin-6 but inhibited that of tumor necrosis factor-α, thereby alleviating liver necrosis and promoting regeneration following CCl₄ injury. The 3D culture of hUC-MSCs thus promotes cell yield and stemness maintenance and represents a promising strategy for hUC-MSCs expansion on an industrial scale with great potential for cell therapy and biotechnology.