For differentiation of human ES or iPS cells to megakaryocytes and platelets.

STEMdiff™ Megakaryocyte Kit is designed for the serum-free and feeder-free differentiation of human embryonic stem (ES) and induced pluripotent stem (iPS) cells to polyploid megakaryocytes capable of shedding platelets and expressing CD41a and CD42b. This simple two-dimensional 17-day protocol includes two stages. During the first 12 days, STEMdiff™ Hematopoietic Supplement A and then Supplement MK1 are added to basal medium to induce cells toward megakaryocyte-biased hematopoietic progenitor cells. At the end of this stage, hematopoietic progenitor cells are easily harvested from the culture supernatant, and are further differentiated to mature megakaryocytes using Supplement MK2 and StemSpan™ SFEM II for an additional 5 days.

At the end of the protocol (Day 17), the cells are typically expanded 405-fold +/- 54-fold (95% CI), and on average 71% +/- 3.3% (95 CI) of cells co-express CD41a and CD42b. STEMdiff™ Megakaryocyte Kit has been optimised for differentiation of ES/iPS cells maintained in mTeSR™1, mTeSR™ Plus, or TeSR™-AOF.