iPSC-derived T Cells, Natural Killer (NK) Cells & Hematopoietic Progenitor Cells (HPC)
Applied StemCell has developed an efficient integration-free, small molecule-based method to
differentiate high-quality T cells, natural killer (NK) cells, and hematopoietic progenitor cells (HPC) from human iPSCs. The differentiated T cells, NKs, and HPCs recapitulate the phenotype and functional parameters of primary and in vivo T cells, NKs. and HPCs. These iPSC-differentiated cells can be used as control lines to compare phenotype and functionality of patient-derived, genome-edited iPSC-derived cells for drug screening applications. These cells can also be used as effector cells for cytotoxicity assays.
- iPSC-derived Natural Killer (NK) Cells
- iPSC-derived T lymphocytes (T cells)
- High Purity CD8+ T Cells Now Available! (Approximately 80-90% purity)
- iPSC-derived Hematopoietic Progenitor Cells (HPC)
- Affordable Prices
- Fast Turn-Around Time
Figure 1: iPSC Differentiation Process.
T lymphocytes (T cells): With the FDA approval of two novel T cell-based therapies for treating cancer, Kymriah™ and Yescarta™, the race for immunotherapies for different types of cancer and immune system-based disorders has intensified. These are a type of adoptive cell transfer (ACT) therapies called chimeric antigen receptor-T cell (CAR-T) where a patient’s (autologous) T cells are genetically engineered ex vivo, to recognize and kill tumor-specific antigens expressed on the surface of tumor cells. They leverage the central role played by T lymphocytes in the immune response to treat cancer. Another type of cytotoxic T lymphocyte cell therapy includes directly delivering ex vivo engineered autologous T cell receptors (TCR-T) that recognize tumor-specific intracellular proteins presented as protein fragments by the major histocompatibility complex (MHC) class I proteins.
Although a great success initially in cancer remission and treatment, there are limitations in the technology such as: limited to autologous (patient) T cells, loss of naïve T cells and persistence of long-term immunologic memory as T cells acquire effector functions, and T cell exhaustion due to repeated activation of T cells in cancer patients which severely reduces the amount of cells that can be harvested from the patient. Such limitations may be overcome by using T cells differentiated ex vivo from iPSCs reprogrammed from patient cells. These T cells have rejuvenated T cell characteristics and can be produced in high numbers with consistent and reliable quality. Additionally, iPSCs can be genetically engineering more efficiently than primary T cells and can be used for generating CAR-T and TCR-T cells for immunotherapy applications.
Natural Killer (NK) cells are a type of lymphoid cells that originate from the same progenitor as T cells and B cells. They are an important part of the innate immune response. NK cells are activated in response to interferons or macrophage-derived cytokines. They recognize “non-self” cells without the need for antigen presentation or recognition, executing a rapid immune reaction. The broad cytotoxicity and rapid apoptosis induced by the NK cells help contain virus-infected cells and controlling early signs of cancer while the adaptive immune response is activated to produce cytotoxic T cells to clear the antigens. With the improving success rate of CAR-T cell therapy, scientists are exploring the NK cells to engineer them for CAR-NK cell therapy to target cancer cells. CAR-NK cells have certain advantages over CAR-T cells in that they retain their natural tumor recognition and killing ability; they do not require strict HLA matching and lack the potential to cause graft-versus-host disease (GvHD), they do not have the same safety concerns as seen with CAR-T cells such as cytokine release syndrome, and therefore they can be generated from allogenic donors and possibly developed as off-shelf therapeutic products.
However, purification of NK cells from allogenic hosts is critical since residual T and B cells may cause GvHD and other complications. Primary NK cells are difficult to harvest, purify, and transfect/transduce, and result in a heterogenous population. They are also difficult to standardize due to the heterogeneity in starting material from different donors. As well, the generation of large quantities of highly pure NK cells requires an extended manufacturing process which can compromise recovery of NK cells, their viability, and potency. Therefore, iPSC-derived NK cells offer a consistent and renewable starting material to provide standardizable NK cells without the dependency on donors and complex harvesting and purification processes. They offer a method to generate homogenous populations of CD56+ CD45+ NK cells and subsequently to generate CAR-NK cells for targeted allogenic cancer immunotherapy, even solid tumors.
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Kawamoto, H., Masuda, K., Nagano, S., & Maeda, T. (2018). Cloning and expansion of antigen-specific T cells using iPS cell technology: development of “off-the-shelf” T cells for the use in allogeneic transfusion settings. International journal of hematology, 107(3), 271-277.
Minagawa, A., Yoshikawa, T., Yasukawa, M., Hotta, A., Kunitomo, M., Iriguchi, S., ... & Kawai, Y. (2018). Enhancing T cell receptor stability in rejuvenated iPSC-derived T cells improves their use in cancer immunotherapy. Cell stem cell, 23(6), 850-858.
Natural killer cells for cancer immunotherapy: a new CAR is catching up. (). EBioMedicine, 39, 1–2. doi:10.1016/j.ebiom.2019.01.018.
Zeng, J., Tang, S. Y., Toh, L. L., & Wang, S. (). Generation of "Off-the-Shelf" Natural Killer Cells from Peripheral Blood Cell-Derived Induced Pluripotent Stem Cells. Stem cell reports, 9(6), 1796–1812. doi:10.1016/j.stemcr.2017.10.020.
Characterization of the ASE-9708 NK Cells
Figure 1. Flow cytometry analysis of ASE-9708 iPSC-derived NK cells for NK cell biomarkers. Cryopreserved NKs, differentiated from Applied StemCell’s control iPSC line, ASE-9211 were recovered in NK culture media. The cells were stained with NK cell markers, CD45 and CD56 at day 2. Left: Isotype control antibodies. Right: CD45/CD56.
Characterization of the ASE-9718 Hematopoietic Progenitor Cells
Figure 2. Flow cytometry analysis of ASE-9718 iPSC-derived Hematopoietic Progenitor Cells for an HPC biomarker. Cryopreserved HPCs, differentiated from Applied StemCell’s control iPSC line, ASE-9211 were recovered in HPC culture basal media. The cells were stained with HPC cell marker, CD34 (right panel) or isotype control (Left panel).
Characterization of the ASE-9719 Hematopoietic Progenitor Cells
Figure 3. Flow cytometry analysis of ASE-9719 iPSC-derived Hematopoietic Progenitor Cells for HPC biomarkers. Cryopreserved HPCs, differentiated from Applied StemCell’s control iPSC line, ASE-9211 were recovered in HPC culture basal media. The cells were stained with HPC markers, Top: CD34 (Right panel) or isotype control (Left panel), Bottom: CD43 (Right panel) or isotype control (Left panel).