Cellular Characterization of Human Pluripotent Stem Cells

  • Rene H. QuintanillaJr.
Part of the Methods in Molecular Biology book series (MIMB, volume 997)


Human pluripotent stem cells (PSCs), in particular induced PSCs, are very difficult to derive, grow, and bank. They require extensive amounts of resources and time to render them useful for basic and applied research. As the derivation methods, culture systems and tissues of origin differ, so does the quality of the PSCs themselves. Consequently, there are generally accepted molecular and cellular markers that serve as benchmarks of pluripotency. PSCs undergo rigorous qualification before they can be truly considered a stem cell or completely reprogrammed into a stem-like cell as in the case of iPSCs. Morphology is a good indicator of PSCs but the further qualification of cellular markers of pluripotency and differential potential is necessary. The standard methods enclosed in this chapter delineate the techniques necessary to qualify PSCs at the cellular level.

Key words

hESC iPSC Immunofluorescence Cellular characterization Human pluripotent stem cells 



This work was supported by Life Technologies. The products within this publication are for Research Use Only, not intended for animal or human therapeutic or diagnostic use.


  1. 1.
    Thomson JA, Itskovitz-Eldor J, Shapiro SS, Waknitz MA, Swiergiel JJ, Marshal VS, Jones JM (1998) Embryonic stem cell lines derived from human blastocysts. Science 282:1145–1147PubMedCrossRefGoogle Scholar
  2. 2.
    Pera MF, Reubinoff B, Trouson A (2000) Human embryonic stem cells. J Cell Sci 113:5–10PubMedGoogle Scholar
  3. 3.
    Takahashi K, Tanabe T, Ohnuki M, Narita M, Ichisaka T, Tomoda K, Yamanaka S (2007) Induction of pluripotent stem cells from adult human fibroblasts by defined factors. Cell 131:861–872PubMedCrossRefGoogle Scholar
  4. 4.
    Josephson R, Ording CJ, Liu Y, Shin S, Lakshmipathy U, Toumadje A, Love B, Chesnut JD, Andrews PW, Rao MS, Auernach JM (2007) Qualification of embryonal carcinoma 2102Ep as a reference for human embryonic stem cell research. Stem Cells 25:437–446PubMedCrossRefGoogle Scholar
  5. 5.
    Chan EM, Ratanasirintrawoot S, Park I, Manos PD, Loh Y, Huo H, Miller JD, Hartung O, Rho J, Ince TA, Daley GQ, Schlaeger TM (2009) Live cell imaging distinguishes bona fide human iPSC cells from partially reprogrammed cells. Nat Biotechnol 27:1033–1037PubMedCrossRefGoogle Scholar
  6. 6.
    Tavakoli T, Xu X, Derby E, Serebryakova Y, Reid Y, Rao MS, Mattson MP, Ma W (2009) Self-renewal and differentiation capabilities are variable between human embryonic stem cell lines 13, 16 and BG01V. BMC Cell Biol 10:44PubMedCrossRefGoogle Scholar
  7. 7.
    Singh U, Quintanilla RH, Grecian S, Gee KR, Rao MS, Lakshmipathy U (2012) Novel live alkaline phosphatase substrate for identification of pluripotent stem cells. Stem Cell Rev 8(3):1021–1029PubMedCrossRefGoogle Scholar
  8. 8.
    Skottman H (2010) Derivation and characterization of three new human embryonic stem cell lines in Finland. In Vitro Cell Dev Biol Anim 46:206–209PubMedCrossRefGoogle Scholar
  9. 9.
    Itskovitz-Eldor J, Schuldiner M, Karsenti D, Eden A, Yanuka O, Amit M, Soreq H, Benvenisty N (2000) Differentiation of human embryonic stem cells into embryoid bodies comprising the three embryonic germ layers. Mol Med 6(2):88–95PubMedGoogle Scholar

Copyright information

© Springer Science+Business Media New York 2013

Authors and Affiliations

  • Rene H. QuintanillaJr.
    • 1
  1. 1.Primary and Stem Cell SystemsLife TechnologiesCarlsbadUSA

Personalised recommendations