Effects of glucose, lactate and basic FGF as limiting factors on the expansion of human induced pluripotent stem cells
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 proces...
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Published in | Journal of bioscience and bioengineering Vol. 125; no. 1; pp. 111 - 115 |
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Main Authors | , , , |
Format | Journal Article |
Language | English |
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Elsevier B.V
01.01.2018
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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. |
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AbstractList | 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. |
Author | Urabe, Yusuke Horiguchi, Ikki Sakai, Yasuyuki Kimura, Keiichi |
Author_xml | – sequence: 1 givenname: Ikki surname: Horiguchi fullname: Horiguchi, Ikki email: ikki.horiguchi@gmail.com organization: Department of Chemical System Engineering, The University of Tokyo, 7-3-1 Hongo, Bunkyo-ku, Tokyo 113-8656, Japan – sequence: 2 givenname: Yusuke surname: Urabe fullname: Urabe, Yusuke organization: Department of Chemical System Engineering, The University of Tokyo, 7-3-1 Hongo, Bunkyo-ku, Tokyo 113-8656, Japan – sequence: 3 givenname: Keiichi surname: Kimura fullname: Kimura, Keiichi organization: Department of Chemical System Engineering, The University of Tokyo, 7-3-1 Hongo, Bunkyo-ku, Tokyo 113-8656, Japan – sequence: 4 givenname: Yasuyuki surname: Sakai fullname: Sakai, Yasuyuki organization: Department of Chemical System Engineering, The University of Tokyo, 7-3-1 Hongo, Bunkyo-ku, Tokyo 113-8656, Japan |
BackLink | https://www.ncbi.nlm.nih.gov/pubmed/28864123$$D View this record in MEDLINE/PubMed |
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Copyright | 2017 The Society for Biotechnology, Japan Copyright © 2017 The Society for Biotechnology, Japan. Published by Elsevier B.V. All rights reserved. |
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Keywords | Heparan sulfate Mass production Lactate Induced pluripotent stem cells Regenerative medicine Cell expansion Basic fibroblast growth factor Glucose |
Language | English |
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Snippet | Pluripotent stem cells (PSCs) are one of the promising cell sources for tissue engineering and drug screening. However, mass production of induced pluripotent... |
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SubjectTerms | Basic fibroblast growth factor Cell Differentiation - drug effects Cell expansion Cell Proliferation - drug effects Culture Media - economics Culture Media - pharmacology Fibroblast Growth Factor 2 - pharmacology Glucose Glucose - metabolism Glucose - pharmacology Heparan sulfate Humans Hydrogen-Ion Concentration Induced pluripotent stem cells Induced Pluripotent Stem Cells - cytology Induced Pluripotent Stem Cells - drug effects Induced Pluripotent Stem Cells - metabolism Lactate Lactic Acid - metabolism Lactic Acid - pharmacology Mass production Regenerative medicine Time Factors |
Title | Effects of glucose, lactate and basic FGF as limiting factors on the expansion of human induced pluripotent stem cells |
URI | https://dx.doi.org/10.1016/j.jbiosc.2017.08.004 https://www.ncbi.nlm.nih.gov/pubmed/28864123 https://search.proquest.com/docview/1935810224 |
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