Vascular Biology Program and Department of Surgery, Children's Hospital Boston, Harvard Medical School, Boston, MA, USA.
Adv Biochem Eng Biotechnol. 2009;112:209-29. doi: 10.1007/978-3-540-69357-4_9.
Expansion of the cell population in vitro has become an essential step in the process of tissue engineering and also the systematic optimization of culture conditions is now a fundamental problem that needs to be addressed. Herein, we provide a rational methodology for searching culture conditions that optimize the acquisition of large quantities of cells following a sequential expansion process. In particular, the analysis of both seeding density and passage length was considered crucial, and their correct selection should be taken as a requisite to establish culture conditions for monolayer systems. This methodology also introduces additional considerations concerning the running cost of the expansion process. The selection of culture conditions will be a compromise between optimal cell expansion and acceptable running cost. This compromise will normally translate into an increase of passage length further away from the optimal value dictated by the growth kinetic of the cells. Finally, the importance of incorporating functional assays to validate the phenotypical and functional characteristics of the expanded cells has been highlighted. The optimization approach presented will contribute to the development of feasible large scale expansion of cells required by the tissue engineering industry.
体外细胞群体扩增已成为组织工程过程中的一个必要步骤，同时，系统优化培养条件现在是一个需要解决的基本问题。在此，我们提供了一种合理的方法来寻找培养条件，以优化大量细胞的获取，遵循一个连续的扩增过程。特别是，分析接种密度和传代长度被认为是至关重要的，它们的正确选择应该被视为建立单层系统培养条件的必要条件。该方法还考虑了扩展过程运行成本的额外因素。培养条件的选择将是在最佳细胞扩增和可接受的运行成本之间的一种妥协。这种妥协通常会导致传代长度的增加，远离细胞生长动力学所规定的最佳值。最后，强调了将功能测定纳入其中以验证扩增细胞的表型和功能特征的重要性。所提出的优化方法将有助于组织工程行业所需的可行的大规模细胞扩增的发展。
