Figure 1. Normal hES and hiPS Cell Morphology is Observed in cGMP mTeSR™1 Cultures
Undifferentiated (A) H1 human embryonic stem (hES) and (B) WLS-1C human induced pluripotent stem (hiPS) cells cultured on Corning® Matrigel® Matrix in cGMP mTeSR™1 retain the prominent nucleoli and high nuclear-to-cytoplasmic ratio characteristic of this cell type after 10 passages. Densely packed cells and multi-layering are prominent when cells are ready to be passaged.
Figure 2. High Expansion Rates are Observed in cGMP mTeSR™1 Cultures
Graph shows the average fold expansion per passage +/- SEM obtained for hES (H1 and H9) and hiPS (WLS-1C) cells cultured in cGMP mTeSR™1 (red) or non-cGMP mTeSR™1 (gray) on Corning® Matrigel® Matrix over 10 passages. Expansion was determined by enumerating the cell aggregates obtained at harvest and dividing by the number of cell aggregates seeded. Note that this data is representative of cultures passaged after 6-7 days in culture, lower expansion should be expected if using shorter culture times.
Figure 3. Cells Cultured in cGMP mTeSR™1 Medium Express Undifferentiated Cell Markers
Histogram analysis for hES (H1 and H9) and hiPS (WLS-1C) cells characterized using FACS for undifferentiated cell markers, OCT4 (OCT3) (Catalog #60093) and TRA-1-60 (Catalog #60064), after 8 - 10 passages in cGMP mTeSR™1 (filled = sample, blank = isotype control).
Figure 4. hPSCs Maintained in cGMP mTeSR™1 Display a Normal Karyotype
Karyograms of (A) H1 hES and (B) WLS-1C hiPS cells cultured in cGMP mTeSR™1 for 11 passages shows that a normal karyotype is retained.
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Applications
This product is designed for use in the following research area(s) as part of the highlighted workflow stage(s). Explore these workflows to learn more about the other products we offer to support each research area.
Conditional CRISPR-mediated deletion of Lyn kinase enhances differentiation and function of iPSC-derived megakaryocytes. A. J. Moroi and P. J. Newman Journal of thrombosis and haemostasis : JTH 2022 jan
Abstract
BACKGROUND Thrombocytopenia leading to life-threatening excessive bleeding can be treated by platelet transfusion. Currently, such treatments are totally dependent on donor-derived platelets. To support future applications in the use of in vitro-derived platelets, we sought to identify genes whose manipulation might improve the efficiency of megakaryocyte production and resulting hemostatic effectiveness. Disruption of Lyn kinase has previously been shown to improve cell survival, megakaryocyte ploidy and TPO-mediated activation in mice, but its role in human megakaryocytes and platelets has not been examined. METHODS To analyze the role of Lyn at defined differentiation stages during human megakaryocyte differentiation, conditional Lyn-deficient cells were generated using CRISPR/Cas9 technology in iPS cells. The efficiency of Lyn-deficient megakaryocytes to differentiate and become activated in response to a range of platelet agonists was analyzed in iPSC-derived megakaryocytes. RESULTS Temporally controlled deletion of Lyn improved the in vitro differentiation of hematopoietic progenitor cells into mature megakaryocytes, as measured by the rate and extent of appearance of CD41+ CD42+ cells. Lyn-deficient megakaryocytes also demonstrated improved hemostatic effectiveness, as reported by their ability to mediate clot formation in rotational thromboelastometry. Finally, Lyn-deficient megakaryocytes produced increased numbers of platelet-like particles (PLP) in vitro. CONCLUSIONS Conditional deletion of Lyn kinase increases the hemostatic effectiveness of megakaryocytes and their progeny as well as improving their yield. Adoption of this system during generation of in vitro-derived platelets may contribute to both their efficiency of production and their ability to support hemostasis.
Establishment of a human iPSC line (SUTCMi001-A) derived from a healthy donor. L. Min et al. Stem cell research 2022 aug
Abstract
This study describes the characterization of one induced pluripotent stem cell line (iPSC) from a healthy female. It is crucial to use iPSCs derived from healthy individuals as controls in genetic disease studies. Thus, we established a human iPSC cell line derived from healthy people. The iPSC cell line was generated in our lab from the peripheral blood mononuclear cells (PBMCs) of a 28-year-old girl. The generated hiPSC line is free of episomal vectors, has a normal karyotype, expresses pluripotency markers and can differentiate into three germ layers in vivo.
iPSC-Based Modeling of RAG2 Severe Combined Immunodeficiency Reveals Multiple T Cell Developmental Arrests. M. Themeli et al. Stem cell reports 2020 feb
Abstract
RAG2 severe combined immune deficiency (RAG2-SCID) is a lethal disorder caused by the absence of functional T and B cells due to a differentiation block. Here, we generated induced pluripotent stem cells (iPSCs) from a RAG2-SCID patient to study the nature of the T cell developmental blockade. We observed a strongly reduced capacity to differentiate at every investigated stage of T cell development, from early CD7-CD5- to CD4+CD8+. The impaired differentiation was accompanied by an increase in CD7-CD56+CD33+ natural killer (NK) cell-like cells. T cell receptor D rearrangements were completely absent in RAG2SCID cells, whereas the rare T cell receptor B rearrangements were likely the result of illegitimate rearrangements. Repair of RAG2 restored the capacity to induce T cell receptor rearrangements, normalized T cell development, and corrected the NK cell-like phenotype. In conclusion, we succeeded in generating an iPSC-based RAG2-SCID model, which enabled the identification of previously unrecognized disorder-related T cell developmental roadblocks.
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Legal Statement: This product was developed under license to intellectual property owned by WiCell™ Research Institute. This product is sold for research use only (whether the buyer is an academic or for-profit entity) under a non-transferable, limited-use license. Purchase of this product does not include the right to sell, use or otherwise transfer this product for commercial purposes (i.e., any activity undertaken for consideration, such as use of this product for manufacturing, or resale of this product or any materials made using this product, or use of this product or any materials made using this product to provide services) or clinical use (i.e., administration of this product or any material using this product to humans) or the right to implant any material made using this product into an animal by, or in collaboration with, a for-profit entity, for purposes other than basic pre-clinical research applications (including without limitation teratoma assays) to validate the function of the cells. Purchasers who do not agree to the terms and conditions set forth above should return the product in acceptable conditions to the seller for a refund. 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.