T Cell, NK Cell Differentiation from Induced Pluripotent Stem Cells

T cells and NK cells are both types of white blood cells that play important roles in the immune system. T cells (T lymphocytes) are responsible for killing infected cells or producing cytokines to activate other immune cells. NK cells (natural killer cells) directly kill infected cells without prior activation, and they also produce cytokines to activate other immune cells.

Fig, 1 Schematic of the overall approach to generate iNK and iT cells from engineered iPSCs. (Cichocki F, et al., 2023)

iPSC-derived T cells, NK cells, dendritic cells, and others provide a next-generation toolset for understanding cancer and disease pathology as well as developing successful immunotherapies (adoptive therapies like CAR-T, CAR-NK, TCR-T, and more).

T Cells

• T cells, a type of white blood cell, play a crucial role in the adaptive immune response. The process of differentiating iPSCs into T cells involves several distinct stages. First, iPSCs are directed toward mesodermal lineage specification through the activation of specific signaling pathways. This is achieved by exposing the iPSCs to growth factors and cytokines known to promote mesoderm development.
• Once mesodermal commitment is achieved, iPSCs are induced to differentiate into hematopoietic progenitor cells (HPCs). This is accomplished through the modulation of key transcription factors and signaling pathways. HPCs are capable of giving rise to multiple blood cell lineages, including T cells. By carefully manipulating the culture conditions and providing appropriate growth factors, iPSC-derived HPCs can be guided toward the T cell lineage.
• Subsequent stages involve the maturation and selection of T cells. This is achieved by mimicking the complex microenvironment of the thymus, where T cells undergo maturation and education. Various co-culture systems and specialized media can be employed to enhance the efficiency and fidelity of T cell differentiation from iPSCs.

NK Cells

• The protocols for generating NK cells from pluripotent stem cells have evolved.
• Human ESCs were cocultured with murine bone marrow stromal cells and then sorted and co-cultured with a second stromal line in media supplemented with cytokines that support NK-cell differentiation.
• This protocol was then modified to adopt a spin-embryoid body method to cultivate hematopoietic progenitor cells in stroma-free, serum-free conditions before differentiation toward the NK-cell lineage. NK cells generated from ESCs exhibited antitumor and antiviral activity both in vitro and in vivo.

Other Lymphocytes

It is worth mentioning that other immune cell types with potential clinical benefits have been generated from iPSCs. γδ T cells have therapeutic potential with a low risk of graft-versus-host-disease, owing to their highly restricted TCR profile. In a two-step process to generate human Vγ9Vδ2 T cells with abundant expression of NK cell–associated activating receptors, primary Vγ9Vδ2 T cells were reprogrammed to iPSCs and then differentiated along the γδ T cell lineage using an NK cell-promoting protocol.

Creative Bioarray Relevant Recommendations

Creative Bioarray is an experienced and outstanding provider of T cell and NK cell differentiation services. With its world-class comprehensive stem cell platform including iPSC generation, genome editing, and differentiation, we provide services to differentiate your iPSCs into CD34+ hematopoietic lineage progenitor cells, and further into high-quality lineage committed immune cells, CD8+ T cells, NK cell, dendritic cells, monocytes, and more.

Reference

1. Cichocki F, et al. (2023). "Engineered and banked iPSCs for advanced NK- and T-cell immunotherapies." Blood. 141 (8), 846-855.