Abstract
This chapter illustrates the patterns observed in situ in populations of pluripotent stem cells (PSCs) when they are stained with fluorescently tagged antibodies to detect antigens expressed on the cell surface or within the cells, thereby identifying a specific molecular marker. These patterns make it possible to judge the homogeneity of a population of cells. A core set of antibodies are used to determine the undifferentiated status of a PSC line. These include SSEA-1, SSEA-3, SSEA-4, Tra-1-60, Tra-1-81 and Oct-4 (octamer-binding transcription factor 4). It is prudent to align the morphology of the colony with the staining pattern to demonstrate concordance.
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Atlasi Y, Mowla SJ, Ziaee SA, Gokhale PJ, Andrews PW. OCT4 spliced variants are differentially expressed in human pluripotent and non-pluripotent cells. Stem Cells. 2008;26:3068–74.
Calloni R, Cordero EA, Henriques JA, Bonatto D. Reviewing and updating the major molecular markers for stem cells. Stem Cells Dev. 2013;22:1455–76.
Draper JS, Pigott C, Thomson JA, Andrews PW. Surface antigens of human embryonic stem cells: changes upon differentiation in culture. J Anat. 2002;200:249–58.
International Stem Cell Initiative, Adewumi O, Aflatoonian B, Ahrlund-Richter L, Amit M, Andrews PW, et al. Characterization of human embryonic stem cell lines by the International Stem Cell Initiative. Nat Biotechnol. 2007;25:803–16.
Kannagi R, Cochran NA, Ishigami F, Hakomori S, Andrews PW, Knowles BB, Solter D. Stage-specific embryonic antigens (SSEA-3 and -4) are epitopes of a unique globo-series ganglioside isolated from human teratocarcinoma cells. EMBO J. 1983;2:2355–61.
Laevsky I. Immunostaining. 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; 2012. p. 105–14.
Nichols J, Zevnik B, Anastassiadis K, Niwa H, Klewe-Nebenius D, Chambers I, et al. Formation of pluripotent stem cells in the mammalian embryo depends on the POU transcription factor Oct4. Cell. 1998;95:379–91.
Nourse JL, Ziaeian B, Palmer T, Schwartz PH, Flanagan LA. Immunocytochemical analysis of human stem cells. In: Loring JF, Peterson SE, editors. Human stem cell manual. 2nd ed. London: Elsevier; 2012. p. 249–70.
Schopperle WM, DeWolf WC. The TRA-1-60 and TRA-1-81 human pluripotent stem cell markers are expressed on podocalyxin in embryonal carcinoma. Stem Cells. 2007;25:723–30.
Zhao W, Ji X, Zhang F, Li L, Ma L. Embryonic stem cell markers. Molecules. 2012;17:6196–236.
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Healy, L., Ruban, L. (2015). Characteristic Staining Patterns of Undifferentiated and Differentiated 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_8
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DOI: https://doi.org/10.1007/978-1-4899-7507-2_8
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