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Effects of glucose, lactate and basic FGF as limiting factors on the expansion of human induced pluripotent stem cells

Author:
Horiguchi, Ikki, Urabe, Yusuke, Kimura, Keiichi, Sakai, Yasuyuki
Source:
Journal of bioscience and bioengineering 2018 v.125 no.1 pp. 111-115
ISSN:
1389-1723
Subject:
cell growth, culture media, developmental stages, drugs, glucose, heparan sulfate, induced pluripotent stem cells, lactic acid, pH, quantitative polymerase chain reaction, reverse transcriptase polymerase chain reaction, screening, tissue engineering
Abstract:
Pluripotent stem cells (PSCs) are one of the promising cell sources for tissue engineering and drug screening. However, mass production of induced pluripotent stem cells (iPSCs) is still developing. Especially, a huge amount of culture medium usage causes expensive cost in the mass production process. In this report, we reduced culture medium usage by extending interval of changing culture medium. In parallel, we also increased glucose concentration and supplied heparan sulfate to avoid depletion of glucose and bFGF, respectively. Reverse transcription-quantitative polymerase chain reaction (RT-qPCR) analyses showed that reducing medium change frequency increased differentiation marker expressions but high glucose concentration downregulated these expressions. In contrast, heparan sulfate did not prevent differentiation marker expressions. According to analyses of growth rate, cell growth with extended medium change interval was decreased in later stage of log growth phase despite the existence of high glucose concentration and heparan sulfate. This result and culturing iPSCs with lactate showed that the accumulation of excreted lactate decreased the growth rate regardless of pH control. Conclusively, these experiments show that adding glucose and removing lactate are important to expand iPSCs with reduced culture medium usage. This knowledge should be useful to design economical iPSC mass production and differentiation system.
Agid:
5813469