Single cell transcriptome sequencing of expanded human hematopoieic stem/progenitor cells cultured in PCL-PVAc-PEG based 3a medium and StemSpan with UM171/SR-1 medium.

We demonstrated that chemically difined PCL-PVAc-PEG based 3a medium was superior to conventional StemSpan besed medium in expansion of human hematopoietic stem cells. In fact, more phenotypic HSCs (CD34+EPCR+CD90+CD45RA-ITGA3+) were expanded in our culture condition and higher donor chmerism was achieved in transplantation assay using humanized mice. But detail transcriptome profiles of expanded HSCs was unknown and therefore single-cell RNA sequencing was done to reveal it. As a result, expanded cells in our medium had more HSPC featured cells. Interestingly, erythroid/megakaryocyte progenitors was more expanded in our condition. On the other hands, other culture condition made more myeloid progenitor cells but few HSPC featured cells. This result support the idea that our new medium are able to selectively expand HSCs. Overall design: Transcriptome analysis of expanded cells at single-cell level to compare efficiency of chemically difined our new medium with conventional StemSpan and cytokine besed medium.