title: Superior Red Blood Cell Generation from Human Pluripotent Stem Cells Through a Novel Microcarrier-Based Embryoid Body Platform.
abstract: In vitro generation of red blood cells (RBCs) from human embryonic stem cells and human induced pluripotent stem cells appears to be a promising alternate approach to circumvent shortages in donor-derived blood supplies for clinical applications. Conventional methods for hematopoietic differentiation of human pluripotent stem cells (hPSC) rely on embryoid body (EB) formation and/or coculture with xenogeneic cell lines. However, most current methods for hPSC expansion and EB formation are not amenable for scale-up to levels required for large-scale RBC generation. Moreover, differentiation methods that rely on xenogenic cell lines would face obstacles for future clinical translation. In this study, we report the development of a serum-free and chemically defined microcarrier-based suspension culture platform for scalable hPSC expansion and EB formation. Improved survival and better quality EBs generated with the microcarrier-based method resulted in significantly improved mesoderm induction and, when combined with hematopoietic differentiation, resulted in at least a 6-fold improvement in hematopoietic precursor expansion, potentially culminating in a 80-fold improvement in the yield of RBC generation compared to a conventional EB-based differentiation method. In addition, we report efficient terminal maturation and generation of mature enucleated RBCs using a coculture system that comprised primary human mesenchymal stromal cells. The microcarrier-based platform could prove to be an appealing strategy for future scale-up of hPSC culture, EB generation, and large-scale generation of RBCs under defined and xeno-free conditions.
authors.affiliation: 1 Stem Cell Group, Bioprocessing Technology Institute , Agency for Science, Technology and Research, Singapore, Republic of Singapore .
authors.forename: Jaichandran
authors.initials: J
authors.lastname: Sivalingam
authors.affiliation: 1 Stem Cell Group, Bioprocessing Technology Institute , Agency for Science, Technology and Research, Singapore, Republic of Singapore .
authors.forename: Alan Tin-Lun
authors.initials: AT
authors.lastname: Lam
authors.affiliation: 2 Institute of Molecular and Cellular Biology , Agency for Science, Technology and Research, Singapore, Republic of Singapore .
authors.forename: Hong Yu
authors.initials: HY
authors.lastname: Chen
authors.affiliation: 2 Institute of Molecular and Cellular Biology , Agency for Science, Technology and Research, Singapore, Republic of Singapore .
authors.forename: Bin Xia
authors.initials: BX
authors.lastname: Yang
authors.affiliation: 1 Stem Cell Group, Bioprocessing Technology Institute , Agency for Science, Technology and Research, Singapore, Republic of Singapore .
authors.forename: Allen Kuan-Liang
authors.initials: AK
authors.lastname: Chen
authors.affiliation: 1 Stem Cell Group, Bioprocessing Technology Institute , Agency for Science, Technology and Research, Singapore, Republic of Singapore .
authors.forename: Shaul
authors.initials: S
authors.lastname: Reuveny
authors.affiliation: 2 Institute of Molecular and Cellular Biology , Agency for Science, Technology and Research, Singapore, Republic of Singapore .|3 Department of Biological Sciences, National University of Singapore , Singapore, Republic of Singapore .
authors.forename: Yuin-Han
authors.initials: YH
authors.lastname: Loh
authors.affiliation: 1 Stem Cell Group, Bioprocessing Technology Institute , Agency for Science, Technology and Research, Singapore, Republic of Singapore .
authors.forename: Steve Kah-Weng
authors.initials: SK
authors.lastname: Oh
mesh_terms: D018929:Cell Culture Techniques / Q000295:instrumentation* / Q000379:methods*; D002454:Cell Differentiation; D002478:Cells, Cultured; D018920:Coculture Techniques; D016895:Culture Media, Serum-Free; D058732:Embryoid Bodies / Q000166:cytology*; D004912:Erythrocytes / Q000166:cytology*; D006801:Humans; D039904:Pluripotent Stem Cells / Q000166:cytology*
chemical_list: D016895:Culture Media, Serum-Free