Publication:
Multilineage differentiation potential of hematoendothelial progenitors derived from human induced pluripotent stem cells

dc.contributor.authorRatchapong Netsrithongen_US
dc.contributor.authorSiriwal Suwanpitaken_US
dc.contributor.authorBootsakorn Boonkaewen_US
dc.contributor.authorKongtana Trakarnsangaen_US
dc.contributor.authorLung Ji Changen_US
dc.contributor.authorChartsiam Tipgomuten_US
dc.contributor.authorChinnavuth Vatanashevanopakornen_US
dc.contributor.authorKovit Pattanapanyasaten_US
dc.contributor.authorMethichit Wattanapanitchen_US
dc.contributor.otherFaculty of Medicine, Siriraj Hospital, Mahidol Universityen_US
dc.contributor.otherShenzhen Geno-Immune Medical Instituteen_US
dc.date.accessioned2020-11-18T07:59:08Z
dc.date.available2020-11-18T07:59:08Z
dc.date.issued2020-12-01en_US
dc.description.abstract© 2020, The Author(s). Background: Human induced pluripotent stem cells (hiPSCs) offer a renewable source of cells for the generation of hematopoietic cells for cell-based therapy, disease modeling, and drug screening. However, current serum/feeder-free differentiation protocols rely on the use of various cytokines, which makes the process very costly or the generation of embryoid bodies (EBs), which are labor-intensive and can cause heterogeneity during differentiation. Here, we report a simple feeder and serum-free monolayer protocol for efficient generation of iPSC-derived multipotent hematoendothelial progenitors (HEPs), which can further differentiate into endothelial and hematopoietic cells including erythroid and T lineages. Methods: Formation of HEPs from iPSCs was initiated by inhibition of GSK3 signaling for 2 days followed by the addition of VEGF and FGF2 for 3 days. The HEPs were further induced toward mature endothelial cells (ECs) in an angiogenic condition and toward T cells by co-culturing with OP9-DL1 feeder cells. Endothelial-to-hematopoietic transition (EHT) of the HEPs was further promoted by supplementation with the TGF-β signaling inhibitor. Erythroid differentiation was performed by culturing the hematopoietic stem/progenitor cells (HSPCs) in a three-stage erythroid liquid culture system. Results: Our protocol significantly enhanced the number of KDR+ CD34+ CD31+ HEPs on day 5 of differentiation. Further culture of HEPs in angiogenic conditions promoted the formation of mature ECs, which expressed CD34, CD31, CD144, vWF, and ICAM-1, and could exhibit the formation of vascular-like network and acetylated low-density lipoprotein (Ac-LDL) uptake. In addition, the HEPs were differentiated into CD8+ T lymphocytes, which could be expanded up to 34-fold upon TCR stimulation. Inhibition of TGF-β signaling at the HEP stage promoted EHT and yielded a large number of HSPCs expressing CD34 and CD43. Upon erythroid differentiation, these HSPCs were expanded up to 40-fold and displayed morphological changes following stages of erythroid development. Conclusion: This protocol offers an efficient and simple approach for the generation of multipotent HEPs and could be adapted to generate desired blood cells in large numbers for applications in basic research including developmental study, disease modeling, and drug screening as well as in regenerative medicine.en_US
dc.identifier.citationStem Cell Research and Therapy. Vol.11, No.1 (2020)en_US
dc.identifier.doi10.1186/s13287-020-01997-wen_US
dc.identifier.issn17576512en_US
dc.identifier.other2-s2.0-85095818441en_US
dc.identifier.urihttps://repository.li.mahidol.ac.th/handle/123456789/59847
dc.rightsMahidol Universityen_US
dc.rights.holderSCOPUSen_US
dc.source.urihttps://www.scopus.com/inward/record.uri?partnerID=HzOxMe3b&scp=85095818441&origin=inwarden_US
dc.subjectBiochemistry, Genetics and Molecular Biologyen_US
dc.subjectMedicineen_US
dc.titleMultilineage differentiation potential of hematoendothelial progenitors derived from human induced pluripotent stem cellsen_US
dc.typeArticleen_US
dspace.entity.typePublication
mu.datasource.scopushttps://www.scopus.com/inward/record.uri?partnerID=HzOxMe3b&scp=85095818441&origin=inwarden_US
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