Abstract
Therapeutic uses of cells differentiated from human pluripotent stem cells (hPSCs), either embryonic stem (ES) cells or induced pluripotent stem cells (iPSCs), are now being tested in clinical trials, and it is likely that this will lead to increased commercial interest in the clinical translation of promising hPSC research. Recent technical advances in the use of defined media and culture substrates have significantly improved both the simplicity and predictability of growing hPSCs, allowing a much more straightforward application of current good manufacturing practices (cGMP) to the culture of these cells. In addition, the adoption of cGMP-compliant techniques in research environments will both improve the replication of results and make the transition of promising investigations to the commercial sector significantly less cumbersome. However, passaging methods for hPSCs are inherently unpredictable and rely on operator experience and expertise. This is problematic for the cell manufacturing process where operator time and process predictability are often determining cost drivers. We have adopted a human iPSC system using defined media and a recombinant substrate that employs cell dissociation with a hypertonic citrate solution which eliminates variability during hPSC cell expansion and provides a simple cGMP-compliant technique for hiPSC cultivation that is appropriate in both research and commercial applications.
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References
Chen G et al (2011) Chemically defined conditions for human iPSC derivation and culture. Nat Methods 8(5):424–429
Nie Y et al (2014) Scalable passaging of adherent human pluripotent stem cells. PLoS One 9(1):e88012
Acknowledgements
The authors acknowledge support from NIH grant U01HL100407 and the University of Minnesota Foundation.
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© 2015 Springer Science+Business Media New York
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Parr, A.M., Walsh, P.J., Truong, V., Dutton, J.R. (2015). cGMP-Compliant Expansion of Human iPSC Cultures as Adherent Monolayers. In: Turksen, K., Nagy, A. (eds) Induced Pluripotent Stem (iPS) Cells. Methods in Molecular Biology, vol 1357. Humana Press, New York, NY. https://doi.org/10.1007/7651_2015_243
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DOI: https://doi.org/10.1007/7651_2015_243
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Publisher Name: Humana Press, New York, NY
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Online ISBN: 978-1-4939-3055-5
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