Laboratoire Réactions et Génie des Procédés, UPR-CNRS 3349, INPL-ENSAIA, 2 avenue de la Forêt de Haye, 54505 Vandœuvre-lès-Nancy, France.
Biotechnol Prog. 2012 May-Jun;28(3):780-7. doi: 10.1002/btpr.1527. Epub 2012 Feb 28.
Mesenchymal stem cells (MSC) are known to be a valuable cell source for tissue engineering and regenerative medicine. However, one of the main limiting steps in their clinical use is the amplification step. MSC expansion on microcarriers has emerged during the last few years, fulfilling the lack of classical T-flasks expansion. Even if the therapeutic potential of MSC as aggregates has been recently highlighted, cell aggregation during expansion has to be avoided. Thus, MSC culture on microcarriers has still to be improved, notably concerning cell aggregation prevention. The aim of this study was to limit cell aggregation during MSC expansion on Cytodex-1®, by evaluating the impact of several culture parameters. First, MSC cultures were performed at different agitation rates (0, 25, and 75 rpm) and different initial cell densities (25 and 50×10(6) cell g(-1) Cytodex-1®). Then, the MSC aggregates were put into contact with additional available surfaces (T-flask, fresh and used Cytodex-1®) at different times (before and after cell aggregation). The results showed that cell aggregation was partly induced by agitation and prevented in static cultures. Moreover, cell aggregation was dependent on cell density and correlated with a decrease in the total cell number. It was however shown that the aggregated organization could be dissociated when in contact with additional surfaces such as T-flasks or fresh Cytodex-1® carriers. Finally, cell aggregation could be successfully limited in spinner flask by adding fresh Cytodex-1® carriers before its onset. Those results indicated that MSC expansion on agitated Cytodex-1® microcarriers could be performed without cell aggregation, avoiding a decrease in total cell number.
间充质干细胞（MSC）是组织工程和再生医学中有价值的细胞来源。然而，其在临床应用中的主要限制步骤之一是扩增步骤。MSC 在微载体上的扩增在过去几年中已经出现，满足了传统 T 瓶扩增的不足。即使 MSC 作为聚集体的治疗潜力最近已经被强调，但是在扩增过程中细胞聚集必须被避免。因此，MSC 在微载体上的培养仍然需要改进，特别是在防止细胞聚集方面。本研究的目的是通过评估几种培养参数来限制 MSC 扩增过程中的细胞聚集。首先，在不同的搅拌速度（0、25 和 75rpm）和不同的初始细胞密度（25 和 50×10(6)个细胞 g(-1) Cytodex-1®）下进行 MSC 培养。然后，将 MSC 聚集体在不同时间（在细胞聚集之前和之后）与其他可用表面（T 瓶、新鲜和使用的 Cytodex-1®）接触。结果表明，细胞聚集部分是由搅拌引起的，在静止培养中可以防止。此外，细胞聚集与细胞密度有关，并且与总细胞数的减少相关。然而，当与 T 瓶或新鲜 Cytodex-1®载体等其他表面接触时，聚集的组织可以被解离。最后，通过在起始前添加新鲜的 Cytodex-1®载体，可以在搅拌瓶中成功地限制细胞聚集。这些结果表明，在搅拌的 Cytodex-1®微载体上进行 MSC 扩增可以在没有细胞聚集的情况下进行，避免总细胞数的减少。
