2011
DOI: 10.1002/stem.681
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Brief Report: Benchmarking Human Pluripotent Stem Cell Markers During Differentiation Into the Three Germ Layers Unveils a Striking Heterogeneity: All Markers Are Not Equal

Abstract: Pluripotent stem cells (PSC) are functionally characterized by their capacity to differentiate into all the cell types from the three germ layers. A wide range of markers, the expression of which is associated with pluripotency, has been used as surrogate evidence of PSC pluripotency, but their respective relevance is poorly documented. Here, we compared by polychromatic flow cytometry the kinetics of loss of expression of eight widely used pluripotency markers (SSEA3, SSEA4, TRA-1-60, TRA-1-81, CD24, OCT4, NA… Show more

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Cited by 43 publications
(38 citation statements)
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References 24 publications
(37 reference statements)
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“…We conclude that MBNL1 and RBFOX2 alternative splicing programs are conserved. To confirm the specificity of alternative splicing changes for late mesoderm differentiation we compared splicing changes for the 15 genes in mouse and human stem cells that we fully differentiated into late ectoderm, endoderm and mesoderm 5 . In this case we noted that 12 of the alternative splicing events (all excluding OSBPL3, CTTN and CD47) shifted in human and mouse mesoderm and very few of these events changed in ectoderm or endoderm (Fig.…”
Section: Resultsmentioning
confidence: 99%
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“…We conclude that MBNL1 and RBFOX2 alternative splicing programs are conserved. To confirm the specificity of alternative splicing changes for late mesoderm differentiation we compared splicing changes for the 15 genes in mouse and human stem cells that we fully differentiated into late ectoderm, endoderm and mesoderm 5 . In this case we noted that 12 of the alternative splicing events (all excluding OSBPL3, CTTN and CD47) shifted in human and mouse mesoderm and very few of these events changed in ectoderm or endoderm (Fig.…”
Section: Resultsmentioning
confidence: 99%
“…In vitro differentiation. For the three germ layer differentiation inductions, H9 hESCs were used and differentiated using the protocol described below and in a previous study 5 .…”
Section: Discussionmentioning
confidence: 99%
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“…It promises practically limitless potential, with considerable scope remaining for the introduction of new more advanced antibody conjugates to address the fundamental questions that are being raised as the regulatory establishment that controls pluripotent cell states is further elucidated. The application of multiparameter flow cytometry in stem cell research is still in its infancy but recent publications (International Stem Cell Initiative 2007;Carpenter et al 2004;Gang et al 2007;De Rosa et al 2003;Prowse et al 2009;Pruszak et al 2007;Ramirez et al 2011) suggest this technique is actively being incorporated into stem cell laboratories worldwide, representing hopefully the first important steps towards ensuring high resolution data-rich experiments become the accepted norm. (i) shows the distribution of the SSEA 1 +ve population at 95% of the isotype control on the left (shown previously in Figure 2) and (ii) on the right shows same population after the isotype control gate has been increased to 99%.…”
Section: Discussionmentioning
confidence: 99%
“…Most crucially, it allows for the capture of multidimensional data (Jansen et al 2008 Henderson et al 2002;Gang et al 2007;De Rossa et al 2003;Prowse et al 2009;Pruszak et al 2007;Ramirez et al 2011) have demonstrated the benefits of this technique but given the strength of the data these have presented, it is predicted that this number will continue to rise.…”
Section: Introductionmentioning
confidence: 99%