Li Yi, Guo Gang, Li Li, Chen Fei, Bao Ji, Shi Yu-Jun, Bu Hong
Laboratory of Pathology, West China Hospital, Sichuan University, Chengdu, 610041, China.
Cell Tissue Res. 2015 May;360(2):297-307. doi: 10.1007/s00441-014-2055-x. Epub 2015 Mar 8.
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.
间充质干细胞（MSC）移植是治疗多种疾病的一种很有前景的方法。然而，传统的单层细胞培养技术导致细胞增殖缓慢且产量不足，无法满足临床需求。三维（3D）培养系统在重建复杂微环境和理解细胞所处条件方面正受到关注。我们的目标是建立一种新型的3D系统，用于在真实的3D微环境中培养人脐带间充质干细胞（hUC-MSCs），但无需消化或传代。原代hUC-MSCs在悬浮摇床系统上的无血清培养基（SFM）中分离并培养。记录细胞的增殖、表型和多能性等特征。检测了3D培养的hUC-MSCs对四氯化碳（CCl4）诱导的小鼠急性肝衰竭的治疗效果。在3D摇床系统中，hUC-MSCs在SFM中形成球体，并保持高活力和活跃增殖。与单层培养相比，3D培养系统在相同体积内产生了更多的hUC-MSCs细胞。球体表达更高水平的干细胞标志物，并表现出更强的多能性。移植到小鼠体内后，3D hUC-MSCs显著促进了干扰素-γ和白细胞介素-6的分泌，但抑制了肿瘤坏死因子-α的分泌，从而减轻了肝坏死并促进了CCl4损伤后的再生。因此，hUC-MSCs的3D培养促进了细胞产量和干性维持，是一种在工业规模上扩大hUC-MSCs的有前景的策略，在细胞治疗和生物技术方面具有巨大潜力。
