title: A 3D microfibrous scaffold for long-term human pluripotent stem cell self-renewal under chemically defined conditions.
abstract: Realizing the potential of human pluripotent stem cell (hPSC)-based therapy requires the development of defined scalable culture systems with efficient expansion, differentiation and isolation protocols. We report an engineered 3D microfiber system that efficiently supports long-term hPSCs self-renewal under chemically defined conditions. The unique feature of this system lies in the application of a 3D ECM-like environment in which cells are embedded, that affords: (i) uniform high cell loading density in individual cell-laden constructs (∼10(7) cells/ml); (ii) quick recovery of encapsulated cells (<10min at 37°C) with excellent preservation of cell viability and 3D multicellular structure; (iii) direct cryopreservation of the encapsulated cells in situ in the microfibers with >17-fold higher cell viability compared to those cultured on Matrigel surface; (iv) long-term hPSC propagation under chemically defined conditions. Four hPSC lines propagated in the microfibrous scaffold for 10 consecutive passages were capable of maintaining an undifferentiated phenotype as demonstrated by the expression of stem cell markers and stable karyotype in vitro and the ability to form derivatives of the three germ layers both in vitro and in vivo. Our 3D microfibrous system has the potential for large-scale cultivation of transplantable hESCs and derivatives for clinical applications.
authors.affiliation: Institute of Bioengineering and Nanotechnology, 31 Biopolis Way, The Nanos, Singapore 138669, Singapore. hflu@ibn.a-star.edu.sg
authors.forename: Hong Fang
authors.initials: HF
authors.lastname: Lu
authors.forename: Karthikeyan
authors.initials: K
authors.lastname: Narayanan
authors.forename: Sze-Xian
authors.initials: SX
authors.lastname: Lim
authors.forename: Shujun
authors.initials: S
authors.lastname: Gao
authors.forename: Meng Fatt
authors.initials: MF
authors.lastname: Leong
authors.forename: Andrew C A
authors.initials: AC
authors.lastname: Wan
mesh_terms: D000464:Alginates / Q000494:pharmacology*; D000818:Animals; D002449:Cell Aggregation / Q000187:drug effects; D002460:Cell Line; D049109:Cell Proliferation / Q000187:drug effects; D048430:Cell Shape / Q000187:drug effects; D018914:Cells, Immobilized / Q000166:cytology / Q000187:drug effects / Q000378:metabolism; D002686:Chitin / Q000494:pharmacology*; D003094:Collagen / Q000494:pharmacology; D015925:Cryopreservation; D004338:Drug Combinations; D053595:Embryonic Stem Cells / Q000166:cytology / Q000187:drug effects / Q000378:metabolism / Q000648:ultrastructure; D020723:Glucuronic Acid / Q000494:pharmacology; D006603:Hexuronic Acids / Q000494:pharmacology; D006801:Humans; D007797:Laminin / Q000494:pharmacology; D051379:Mice; D016513:Mice, SCID; D010641:Phenotype; D039904:Pluripotent Stem Cells / Q000166:cytology* / Q000187:drug effects / Q000378:metabolism; D011509:Proteoglycans / Q000494:pharmacology; D013724:Teratoma / Q000473:pathology; D013997:Time Factors; D054457:Tissue Scaffolds / Q000737:chemistry*
chemical_list: D000464:Alginates; D004338:Drug Combinations; D006603:Hexuronic Acids; D007797:Laminin; D011509:Proteoglycans; C059485:matrigel; D002686:Chitin; D020723:Glucuronic Acid; D003094:Collagen