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Embryonic Stem Cells
Pluripotent 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, naïve hESCs isolation has been achieved through 1) induced expression of pluripotency factors and growth factors or 2) following incubation with specific combinations of small modulating molecules and growth factors.
ES like Cells: Induced Pluripotent Stem Cells (iPSCs)
iPSCs originate from somatic cells which are reprogrammed to a pluripotent state by the re-expression of key pluripotency factors. Reprogramming of somatic cells was first demonstrated by Takahashi and Yamanaka in 2006, and was achieved through the induction of Oct4, Sox2, Klf4 and c-Myc expression in mouse fibroblasts. Since then, there has been a significant expansion in the range of adult somatic cells reprogrammed to a pluripotent state (e.g., human peripheral blood mononuclear cells, hematopoietic progenitors, hepatocytes, keratinocytes, muscle cells, and adipocyte cells) and in the efficiency of the reprogramming methodologies.
How is Pluripotency Confirmed?
ESC pluripotency in vitro and in vivo may be confirmed through various approaches.
JOY6 hiPSCs were differentiated towards ectoderm, mesoderm, and endoderm fate using Human Pluripotent Stem Cell Functional Identification Kit (R&D Systems, Catalog # SC027B). Flow cytometry, based on the protocol provided in the kit, was used for differentiation analysis. (orange) Differentiated cells have increased expression of lineage-specific markers for ectoderm - Otx2, mesoderm - Brachyury, or endoderm - SOX17 when compared to (blue) undifferentiated JOY6 hiPSCs.
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.
Embryonic Stem Cell Surface Markers
Pluripotent 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.
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