Efficient generation, purification, and expansion of CD34+ hematopoietic progenitor cells from nonhuman primate induced pluripotent stem cells.

Publication Type:

Journal Article

Source:

Blood (2012)

Keywords:

August 2012, Center-Authored Paper, Clinical Research Division

Abstract:

Induced pluripotent stem cell (iPSC) therapeutics are a promising treatment for hematologic genetic and infectious diseases. To assess engraftment, risk of neoplastic formation, and therapeutic benefit in an autologous setting, testing iPSC therapeutics in an appropriate model, such as the pigtail macaque (Macaca nemestrina), is crucial. Here, we developed a chemically defined, scalable and reproducible specification protocol with Bone Morphogenetic Protein 4 (BMP4), Prostaglandin E2 (PGE2), and StemRegenin1 (SR1) for hematopoietic differentiation of Macaca nemestrina (Mn)iPSCs. Sequential co-culture with BMP4, PGE2, and SR1 led to robust MniPSC hematopoietic progenitor cell (HPC) formation. The combination of PGE2 and SR1 increased CD34(+)CD38(-)Thy1(+)CD49f(+)CD45RA(-) cell yield by 6-fold. CD34(+)CD38(-)Thy1(+)CD49f(+)CD45RA(-) cells isolated based on CD34 expression and cultured in SR1 expanded 3-fold and maintained this LT-HSC phenotype. Purified CD34(high) cells exhibited 4-fold higher hematopoietic colony forming potential compared to unsorted HPCs, and had bi-lineage differentiation potential. Based on these studies, we calculated the cell yields that must be achieved at each stage in order to meet a threshold CD34(+) cell dose that is required for engraftment in the pigtail macaque. Our chemically defined protocol will support scale-up and testing of iPSC-derived CD34(high) cell therapies in a clinically relevant nonhuman primate model.