The MC-813-70 antibody reacts with stage-specific embryonic antigen-4 (SSEA-4), a glycolipid carbohydrate antigen expressed on the surface of human embryonal carcinoma (EC), embryonic germ (EG), undifferentiated embryonic stem (ES) and induced pluripotent stem (iPS) cells, a subset of mesenchymal stem cells, and rhesus monkey ES cell lines. No immunoreactivity is evident with undifferentiated mouse EC, EG, ES, and iPS cells. Expression of SSEA-4 is down-regulated following differentiation of human EC, ES, and iPS cells. In contrast, the differentiation of mouse EC, ES, or iPS cells may be accompanied by an increase in SSEA-4 expression.
This antibody clone has been verified for labeling human ES and iPS cells grown in mTeSR™1 (Catalog #85850), and TeSR™2 (Catalog #05860), and has been verified for purity assessments of cells isolated with EasySep™ kits, including EasySep™ Human ES/iPS Cell TRA-1-60 Positive Selection Kit (Catalog #18166) and EasySep™ hESC/hiPSC SSEA-4 Positive Selection Kit (Catalog #18165; partial blocking may be observed).
This antibody clone has been verified for labeling human ES and iPS cells grown in mTeSR™1 (Catalog #85850), and TeSR™2 (Catalog #05860), and has been verified for purity assessments of cells isolated with EasySep™ kits, including EasySep™ Human ES/iPS Cell TRA-1-60 Positive Selection Kit (Catalog #18166) and EasySep™ hESC/hiPSC SSEA-4 Positive Selection Kit (Catalog #18165; partial blocking may be observed).
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Protocols and Documentation
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Resources and Publications
Educational Materials (3)
Publications (6)
Intrinsic Immunity Shapes Viral Resistance of Stem Cells. Cell 2018 JAN
Abstract
Stem cells are highly resistant to viral infection compared to their differentiated progeny; however, the mechanism is mysterious. Here, we analyzed gene expression in mammalian stem cells and cells at various stages of differentiation. We find that, conserved across species, stem cells express a subset of genes previously classified as interferon (IFN) stimulated genes (ISGs) but that expression is intrinsic, as stem cells are refractory to interferon. This intrinsic ISG expression varies in a cell-type-specific manner, and many ISGs decrease upon differentiation, at which time cells become IFN responsive, allowing induction of a broad spectrum of ISGs by IFN signaling. Importantly, we show that intrinsically expressed ISGs protect stem cells against viral infection. We demonstrate the in vivo importance of intrinsic ISG expression for protecting stem cells and their differentiation potential during viral infection. These findings have intriguing implications for understanding stem cell biology and the evolution of pathogen resistance.
Simple suspension culture system of human iPS cells maintaining their pluripotency for cardiac cell sheet engineering. Journal of Tissue Engineering and Regenerative Medicine 2015 DEC
Abstract
In this study, a simple three-dimensional (3D) suspension culture method for the expansion and cardiac differentiation of human induced pluripotent stem cells (hiPSCs) is reported. The culture methods were easily adapted from two-dimensional (2D) to 3D culture without any additional manipulations. When hiPSCs were directly applied to 3D culture from 2D in a single-cell suspension, only a few aggregated cells were observed. However, after 3 days, culture of the small hiPSC aggregates in a spinner flask at the optimal agitation rate created aggregates which were capable of cell passages from the single-cell suspension. Cell numbers increased to approximately 10-fold after 12 days of culture. The undifferentiated state of expanded hiPSCs was confirmed by flow cytometry, immunocytochemistry and quantitative RT-PCR, and the hiPSCs differentiated into three germ layers. When the hiPSCs were subsequently cultured in a flask using cardiac differentiation medium, expression of cardiac cell-specific genes and beating cardiomyocytes were observed. Furthermore, the culture of hiPSCs on Matrigel-coated dishes with serum-free medium containing activin A, BMP4 and FGF-2 enabled it to generate robust spontaneous beating cardiomyocytes and these cells expressed several cardiac cell-related genes, including HCN4, MLC-2a and MLC-2v. This suggests that the expanded hiPSCs might maintain the potential to differentiate into several types of cardiomyocytes, including pacemakers. Moreover, when cardiac cell sheets were fabricated using differentiated cardiomyocytes, they beat spontaneously and synchronously, indicating electrically communicative tissue. This simple culture system might enable the generation of sufficient amounts of beating cardiomyocytes for use in cardiac regenerative medicine and tissue engineering.
OCT4 as a target of miR-34a stimulates p63 but inhibits p53 to promote human cell transformation Cell death & disease 2014 JAN
Abstract
Human cell transformation is a key step for oncogenic development, which involves multiple pathways; however, the mechanism remains unclear. To test our hypothesis whether cell oncogenic transformation shares some mechanisms with the process of reprogramming non-stem cells to induced pluripotent stem cells (iPSC), we studied the relationship among the key factors for promoting or inhibiting iPSC in radiation-transformed human epithelial cell lines derived from different tissues (lung, breast and colon). We unexpectedly found that p63 and OCT4 were highly expressed (accompanied by low expressed p53 and miR-34a) in all transformed cell lines examined when compared with their non-transformed counterparts. We further elucidated the relationship of these factors: the 3p strand of miR-34a directly targeted OCT4 by binding to the 3′ untranslated region (3′-UTR) of OCT4 and, OCT4, in turn, stimulated p63 but inhibited p53 expression by binding to a specific region of the p63 or p53 promoter. Moreover, we revealed that the effects of OCT4 on promoting cell oncogenic transformation were by affecting p63 and p53. These results support that a positive loop exists in human cells: OCT4 upregulation as a consequence of inhibition of miR-34a, promotes p63 but suppresses p53 expression, which further stimulates OCT4 upregulation by downregulating miR-34a. This functional loop contributes significantly to cell transformation and, most likely, also to the iPSC process.
Legal Statement: Alexa Fluor is a registered trademark of Life Technologies Corporation. Antibodies conjugated to Alexa Fluor® are licensed for internal research use only and sale is expressly conditioned on the buyer not using the antibody for manufacturing, performing a service or medical test, or otherwise generating revenue. For use other than research, contact Life Technologies Corporation, 5791 Van Allen Way, Carlsbad, CA 92008 USA or outlicensing@lifetech.com. PRODUCTS ARE FOR RESEARCH USE ONLY AND NOT INTENDED FOR HUMAN OR ANIMAL DIAGNOSTIC OR THERAPEUTIC USES UNLESS OTHERWISE STATED. FOR ADDITIONAL INFORMATION ON QUALITY AT STEMCELL, REFER TO WWW.STEMCELL.COM/COMPLIANCE.
