Growth Factors for Induced Pluripotent Stem Cell Culturing

iPSC-T Cell

The classical approach to generate iPSC-derived T cells starts from:

1. Co-culturing of iPSCs derived from healthy donors with murine bone marrow stromal cell line (C3H10T1/2 or OP9) to generate CD34+ hematopoietic progenitor cells.

2. Hematoproietic progenitors are enriched and co-cultured with OP9 cell line overexpressing DLL-1/DLL-4, with additional cytokines to drive T cell differentiation.

iPSC-NK Cell

To generate iPSC-derived NK cells:

1. The iPSCs derived from healthy donors are co-cultured with the murine bone marrow stromal cell line M210-B4 for ~20 days to allow for the generation of CD34 + hematopoietic progenitors.

2. CD34 + hematopoietic progenitors are then enriched and co-cultured with a monolayer of murine AFT024 (a fetal liver-derived stromal cell line) or EL08-1D2 stroma cells with defined cytokines for 4 to 5 weeks, eventually generating mature NK cells.

iPSC-Macrophage

Formation of iPSC-derived macrophages Mφs requires a more complex methodology:

1. Embryoid body(EBs) formation is the most commonly used method for differentiating iPSC-derived Mφs, utilizing specific modifications such as rotary suspension, microwell approaches, microfluidic hanging drop chips, and others.

2. After formation of iPSC-derived EBs, myeloid Mφs are further induced by supplementation of cytokines, including GM-CSF, M-CSF, IL-3, and IL-4.