STEMdiff Hematopoietic - EB Basal Medium

From STEMCELL Technologies, this basal medium is a foundational component of the STEMdiff hematopoietic workflow used to differentiate human pluripotent stem cells (PSCs) into CD34+ hematopoietic progenitor cells under serum-free, feeder-free culture conditions. Combined with STEMdiff Hematopoietic - EB Supplement A and supplemented with Y-27632, the basal medium is used to prepare EB Formation Medium and EB Medium A, which drive mesoderm specification in AggreWell plates during the first three days of culture. When mixed with STEMdiff Hematopoietic - EB Supplement B, it forms EB Medium B, which supports hematopoietic lineage specification through day 12 of the protocol.

The basal medium is supplied as a shared component of both the STEMdiff NK Cell Kit and the STEMdiff T Cell Kit, where it underpins a four-stage protocol for differentiating PSCs into CD56+ natural killer cells or CD4+CD8+ double-positive T cells without the use of stromal cells. It has also been adapted for scalable suspension culture workflows (2 - 500 mL) using orbital-shaken plates and bottles or PBS-MINI bioreactor vessels, supporting research into early hematopoiesis, immune cell biology, and the development of adoptive immunotherapies.

In the validated protocol, the average frequency of viable CD34+ cells on day 12 across two ES and three iPS cell lines ranged between 31% and 42%, with average yields per well of an AggreWell 400 plate between 3.3 x 10^5 and 7.3 x 10^5 CD34+ cells. Downstream, approximately 210 NK cells or 60 DP T cells can be generated per input PSC-derived CD34+ cell.