Mapping the biogenesis of forward programmed megakaryocytes from induced pluripotent stem cells


Type
Article
Change log
Authors
Abstract

Platelet deficiency, known as thrombocytopenia, can cause haemorrhage and is treated with platelet transfusions. We developed a system for the production of platelet precursor cells, megakaryocytes, from pluripotent stem cells. These cultures can be maintained for >100 days, implying culture renewal by megakaryocyte progenitors (MKPs). However, it is unclear whether the MKP state in vitro mirrors the state in vivo, and MKPs cannot be purified using conventional surface markers. We performed single cell RNA sequencing throughout in vitro differentiation and mapped each state to its equivalent in vivo. This enabled the identification of 5 surface markers which reproducibly purify MKPs, allowing us an insight into their transcriptional and epigenetic profiles. Finally, we performed culture optimisation, increasing MKP production. Altogether, this study has mapped parallels between the MKP states in vivo and in vitro and allowed the purification of MKPs, accelerating the progress of in vitro-derived transfusion products towards the clinic.

Description
Keywords
CambridgeStemCellInstitute
Journal Title
Science Advances
Conference Name
Journal ISSN
2375-2548
2375-2548
Volume Title
8
Publisher
American Association for the Advancement of Science
Sponsorship
Wellcome Trust (203151/A/16/Z)
Wellcome Trust (203151/Z/16/Z)
Wellcome Trust (102160/B/13/Z)
Medical Research Council (MR/R015724/1)
British Heart Foundation (None)
Medical Research Council (MC_PC_17230)