STEMdiff Erythroid Kit

From STEMCELL Technologies, this differentiation system is designed to generate erythroid progenitor cells (erythroblasts) from human ES or iPS cells using a 24-day, two-stage protocol. The workflow is serum- and feeder-free, offering consistent, reproducible performance across multiple hPSC lines. Erythroblasts produced may be used in disease modeling, drug screening, and research into the regulation of erythropoiesis.

The kit comprises two culture media and three supplements: STEMdiff Hematopoietic Basal Medium, StemSpan SFEM II, STEMdiff Hematopoietic Supplement A (for mesodermal specification), STEMdiff Erythroid Supplement E1 (for hematoendothelial and hematopoietic specification), and STEMdiff Erythroid Supplement E2 (for expansion and differentiation of hematopoietic progenitor cells into erythroblasts). The protocol is optimized for hPSCs maintained in mTeSR1, mTeSR Plus, or TeSR-E8 media on Corning Matrigel-coated plates. In the first stage, hPSC aggregates are directed through hematopoietic specification, yielding CD34+CD43+ hematopoietic progenitor cells. In the second stage, these progenitors are replated and driven toward an erythroid fate.

Using the recommended protocol, cell yields may reach 100,000 erythroblasts per input hPSC in 24 days, with typically more than 70% of the resulting cells identified as CD71+GlyA+ erythroblasts. The differentiated cells express a mix of primitive (embryonic) and definitive (fetal, adult) hemoglobin and display typical basophilic erythroblast morphology. An optional 7-day maturation protocol using StemSpan SFEM II supplemented with 3 U/mL recombinant human EPO and 3% human AB serum enables further maturation into CD71low/GlyA+ normoblasts.