Abstract
The maintenance and expansion of undifferentiated pluripotent stem cells (PSCs) in cell culture systems requires the dissociation of cells and their transfer from culture vessel to culture vessel, in a process known as passaging. This chapter includes images acquired from different techniques used to passage PSCs. It is important to minimise the stress to the cells caused by the passaging activity, as this process can promote apoptosis, spontaneous differentiation and culture adaptation. Culture adaptation through selective pressure of cells in culture results in genetically abnormal karyotypes (Chap. 10).
Access this chapter
Tax calculation will be finalised at checkout
Purchases are for personal use only
Suggested Reading
Amit M, Itskovitz-Eldor J. Morphology of human embryonic and induced pluripotent stem cell colonies cultured with feeders. In: Amit M, Itskovitz-Eldor J, editors. Atlas of human pluripotent stem cells: derivation and culturing, stem cell biology and regenerative medicine. New York: Humana Press; 2012a. p. 15–39.
Amit M, Itskovitz-Eldor J. Morphology of human embryonic stem cells and induced pluripotent stem cell colonies cultured in feeder layer-free conditions. In: Amit M, Itskovitz-Eldor J, editors. Atlas of human pluripotent stem cells: derivation and culturing, Stem cell biology and regenerative medicine. New York: Humana Press; 2012b. p. 41–55.
Bajpai R, Lesperance J, Kim M, Terskikh AV. Efficient propagation of single cells Accutase-dissociated human embryonic stem cells. Mol Reprod Dev. 2008;75:818–27.
Beers J, Gulbranson DR, George N, Siniscalchi LI, Jones J, Thomson JA, Chen G. Passaging and colony expansion of human pluripotent stem cells by enzyme-free dissociation in chemically defined culture conditions. Nat Protoc. 2012;7:2029–40.
Costa M, Sourris K, Hatzistavrou T, Elefanty AG, Stanley EG. Expansion of human embryonic stem cells in vitro. Curr Protoc Stem Cell Biol. 2008;5:1C.1.1–1C.1.7.
Ellerström C, Hyllner J, Strehl R. Single cell enzymatic dissociation of human embryonic stem cells: a straightforward, robust, and standardized culture method. Methods Mol Biol. 2010;584:121–34.
Ellerström C, Strehl R, Noaksson K, Hyllner J, Semb H. Facilitated expansion of human embryonic stem cells by single-cell enzymatic dissociation. Stem Cells. 2007;25:1690–6.
Hartung O, Huo H, Daley GQ, Schlaeger TM. Clump passaging and expansion of human embryonic and induced pluripotent stem cells on mouse embryonic fibroblast feeder cells. Curr Protoc Stem Cell Biol. 2010;14:1C.10.1–15.
Heng BC, Liu H, Ge Z, Cao T. Mechanical dissociation of human embryonic stem cell colonies by manual scraping after collagenase treatment is much more detrimental to cellular viability than is trypsinization with gentle pipetting. Biotechnol Appl Biochem. 2007;47:33–7.
Leonardo TR, Schell JP, Nickey KS, Tran HT. Culturing human pluripotent stem cells on a feeder layer. In: Loring JF, Peterson SE, editors. Human stem cell manual. 2nd ed. London: Elsevier; 2012. p. 3–14.
Nie Y, Walsh P, Clarke DL, Rowley JA, Fellner T. Scalable passaging of adherent human pluripotent stem cells. PLoS One. 2014;9:e88012.
Oh SK, Kim HS, Park YB, Seol HW, Kim YY, Cho MS, et al. Methods for expansion of human embryonic stem cells. Stem Cells. 2005;23:605–9.
Ohnuma K, Fujiki A, Yanagihara K, Tachikawa S, Hayashi Y, Ito Y, et al. Enzyme-free passage of human pluripotent stem cells by controlling divalent cations. Sci Rep. 2014;4:4646.
Street CA, Bryan BA. Rho kinase proteins–pleiotropic modulators of cell survival and apoptosis. Anticancer Res. 2011;31:3645–57.
T’joen V, Declercq H, Cornelissen M. Expansion of human embryonic stem cells: a comparative study. Cell Prolif. 2011;44:462–76.
Author information
Authors and Affiliations
Rights and permissions
Copyright information
© 2015 Springer Science+Business Media New York
About this chapter
Cite this chapter
Healy, L., Ruban, L. (2015). Passaging Pluripotent Stem Cells. In: Atlas of Human Pluripotent Stem Cells in Culture. Springer, Boston, MA. https://doi.org/10.1007/978-1-4899-7507-2_7
Download citation
DOI: https://doi.org/10.1007/978-1-4899-7507-2_7
Published:
Publisher Name: Springer, Boston, MA
Print ISBN: 978-1-4899-7506-5
Online ISBN: 978-1-4899-7507-2
eBook Packages: Biomedical and Life SciencesBiomedical and Life Sciences (R0)