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Embryonic Stem CellsPluripotent embryonic stem cells (ESCs) correspond to cells within a developing embryo that have the capacity to generate all the embryonic germ layers (i.e., endoderm, mesoderm and ectoderm), and are able to give rise to all cell types in the body. ESCs may be derived from developing embryos at the pre-implantation blastocyst stage, and specifically from cells within the inner cell mass (ICM). In mice, the pluripotent state of ICM cells (mESCs) is often referred to as a “naïve” state. Following blastocyst implantation, ICM derived cells or mouse epiblast stem cells (mEpiSCs) retain self-renewal capacity but are in a “primed” state of pluripotency. Similarly, conventional ICM derived human ESCs (hESCs) are characterized by “primed” pluripotent properties, a state that is thought to occur in association with the in vitro conditions used for their derivation and maintenance. However, cultivation of naïve hESCs has been achieved through 1) induced expression of pluripotency factors or 2) incubation with specific combinations of small modulating molecules and growth factors.
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How is Pluripotency Confirmed?ESC pluripotency in vitro and in vivo may be confirmed through various approaches.
What is the Difference between Naïve and Primed ESCs?Naïve and primed ESCs share the same capacity for self-renewal and ability to give rise to the three embryonic germ layers. However, only naïve ESCs generate germline chimeras. The expression of specific pluripotency factors differs between stem cells in a naïve vs primed state.
This table contains only a subset of upregulated naïve and primed cell markers. See Ghimire et al. 2018 (mESCs) and Messmer et al. 2019 (hESCs) for a more complete list. Embryonic Stem Cell Surface MarkersPluripotent stem cells express a variety of cell surface proteins which may be used for their characterization and isolation from heterogeneous populations under culture conditions. The combined use of pluripotency markers to identify ESCs facilitates downstream applications aimed at the propagation of ESCs or their differentiation towards specific lineages.
H/M Pluripotent Stem Cell Flow Cytometry Kit [FMC001-NOV] - Mouse D3 embryonic stem cells were stained using reagents included in the Human/Mouse Embryonic Stem Cell Multi-Color Flow Cytometry Kit. Cells were analyzed for expression of pluripotent markers including SSEA-1, SSEA-4, Oct-3/4, and SOX2 by flow cytometry. A. Flow cytometric analysis shows that 91.1% of mouse D3 embryonic stem cells are positive for both Oct-3/4 and SSEA1 expression. B. Flow cytometric analysis data shows that 82.6% of mouse D3 embryonic stem cells are positive for SSEA-1 and negative for SSEA-4 a phenotype consistent with mouse embryonic stem cells. C. Flow cytometric analysis shows that mouse D3 embryonic stem cells express the pluripotent marker SOX2.
H/M Pluripotent Stem Cell Flow Cytometry Kit [FMC001-NOV] - BG01V human embryonic stem cells were stained using reagents included in the Human/Mouse Pluripotent Stem Cell Multi-Color Flow Cytometry Kit. Cells were simultaneously analyzed for expression of pluripotent markers including SSEA-1, SSEA-4, Oct-3/4, and SOX2 by flow cytometry. A. Flow cytometry data shows that 91.9% of BG01V human embryonic stem cells are positive for both Oct-3/4 and SSEA4 expression. B. Flow cytometry data shows that 88.5% of BG01V human embryonic stem cells are positive for SSEA-4 and negative for SSEA-1, a phenotype consistent with human embryonic stem cells. C. Flow cytometric analysis shows that BG01V human embryonic stem cells express the pluripotent marker SOX2. New ExCellerate™ iPSC Expansion MediumSupports robust expansion and maintenance of pluripotent stem cell culture for enhanced consistency and reproducibility.
Select References Bieberich, E., & Wang, G. (2013). Molecular Mechanisms Underlying Pluripotency. In Pluripotent Stem Cells. https://doi.org/10.5772/55596 Boroviak, T., & Nichols, J. (2017). Primate embryogenesis predicts the hallmarks of human naïve pluripotency. Development (Cambridge). https://doi.org/10.1242/dev.145177 Carstea, A. C. (2009). Germline competence of mouse ES and iPS cell lines: Chimera technologies and genetic background. World Journal of Stem Cells. https://doi.org/10.4252/wjsc.v1.i1.22 Ghimire, S., Van Der Jeught, M., Neupane, J., Roost, M. S., Anckaert, J., Popovic, M., … Heindryckx, B. (2018). Comparative analysis of naive, primed and ground state pluripotency in mouse embryonic stem cells originating from the same genetic background. Scientific Reports. https://doi.org/10.1038/s41598-018-24051-5 Kim, J. S., Choi, H. W., Choi, S., & Do, J. T. (2011). Reprogrammed pluripotent stem cells from somatic cells. International Journal of Stem Cells. https://doi.org/10.15283/ijsc.2011.4.1.1 Kumari, D. (2016). States of Pluripotency: Naïve and Primed Pluripotent Stem Cells. In Pluripotent Stem Cells - From the Bench to the Clinic. https://doi.org/10.5772/63202 Messmer, T., von Meyenn, F., Savino, A., Santos, F., Mohammed, H., Lun, A. T. L., … Reik, W. (2019). Transcriptional Heterogeneity in Naive and Primed Human Pluripotent Stem Cells at Single-Cell Resolution. Cell Reports. https://doi.org/10.1016/j.celrep.2018.12.099 Nichols, J., & Smith, A. (2009). Naive and Primed Pluripotent States. Cell Stem Cell. https://doi.org/10.1016/j.stem.2009.05.015 Trusler, O., Huang, Z., Goodwin, J., & Laslett, A. L. (2018). Cell surface markers for the identification and study of human naive pluripotent stem cells. Stem Cell Research. https://doi.org/10.1016/j.scr.2017.11.017 Weinberger, L., Ayyash, M., Novershtern, N., & Hanna, J. H. (2016). Dynamic stem cell states: Naive to primed pluripotency in rodents and humans. Nature Reviews Molecular Cell Biology. https://doi.org/10.1038/nrm.2015.28 Zhao, W., Ji, X., Zhang, F., Li, L., & Ma, L. (2012). Embryonic Stem Cell Markers. Molecules. https://doi.org/10.3390/molecules17066196 |
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