STEMdiff Hematopoietic - EB Supplement B

This animal component-free culture supplement from STEMCELL Technologies is designed for hematopoietic differentiation with optimized lymphoid potential. It is combined with STEMdiff™ Hematopoietic - EB Basal Medium to prepare EB Medium B, which is used during the second stage of a 12-day protocol for differentiating human pluripotent stem cells (PSCs) into CD34+ hematopoietic progenitor cells. After an initial 3-day mesoderm formation stage in AggreWell™ plates, EB Medium B is applied from day 3 through day 12 to promote hematopoietic lineage specification, with embryoid bodies being transferred to non-tissue culture-treated plates on day 5.

The supplement is a component of the STEMdiff™ NK Cell Kit and STEMdiff™ T Cell Kit, supporting the generation of CD34+ cells that can be further differentiated into natural killer (NK) cells and T cells. Across two ES cell lines (H1 and H9) and three iPS cell lines (WLS-1C, STiPS-M001, and STiPS-F016), the average frequency of viable CD34+ cells on day 12 ranged between 31% and 42%, with average yields per well of a 6-well AggreWell™400 plate between 3.3 x 10^5 and 7.3 x 10^5 cells. This hematopoietic differentiation workflow provides a foundation for downstream lymphoid applications, including the production of CD56+ NK cells and CD4+CD8+ double-positive T cells from PSC-derived CD34+ cells.