Culture medium refinement by dialysis for the expansion of human induced pluripotent stem cells in suspension culture

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• 作者：Suman Chandra Nath ; Eiji Nagamori ; Masanobu Horie…
• 关键词：Refining method ; Culture medium ; Dialysis ; hiPSC expansion ; Autocrine factors ; Lactic acid
• 刊名：Bioprocess and Biosystems Engineering
• 出版年：2017
• 出版时间：January 2017
• 年：2017
• 卷：40
• 期：1
• 页码：123-131
• 全文大小：
• 刊物类别：Chemistry and Materials Science
• 刊物主题：Biotechnology; Industrial and Production Engineering; Environmental Engineering/Biotechnology; Industrial Chemistry/Chemical Engineering; Food Science;
• 出版者：Springer Berlin Heidelberg
• ISSN：1615-7605
• 卷排序：40

文摘

Human induced pluripotent stem cells (hiPSCs) secrete essential autocrine factors that are removed along with toxic metabolites when the growth medium is exchanged daily. In this study, after determining the minimum inhibitory level of lactic acid for hiPSCs, a medium refining system was constructed by which toxic metabolites were removed from used culture medium and autocrine factors as well as other growth factors were recycled. Specifically, about 87 % of the basic fibroblast growth factor and 80 % of transforming growth factor beta 1 were retained in the refined medium after dialysis. The refined medium efficiently potentiated the proliferation of hiPS cells in adherent culture. When the refining system was used to refresh medium in suspension culture, a final cell density of (1.1 ± 0.1) × 106 cells mL−1 was obtained, with 99.5 ± 0.2 % OCT 3/4 and 78.3 ± 1.1 % TRA-1-60 expression, on day 4 of culture. These levels of expression were similar to those observed in the conventional suspension culture. With this method, culture medium refinement by dialysis was established to remove toxic metabolites, recycle autocrine factors as well as other growth factors, and reduce the use of macromolecules for the expansion of hiPSCs in suspension culture.