title: The use of nanofibrillar cellulose hydrogel as a flexible three-dimensional model to culture human pluripotent stem cells.
abstract: Human embryonic stem cells and induced pluripotent stem cells have great potential in research and therapies. The current in vitro culture systems for human pluripotent stem cells (hPSCs) do not mimic the three-dimensional (3D) in vivo stem cell niche that transiently supports stem cell proliferation and is subject to changes which facilitate subsequent differentiation during development. Here, we demonstrate, for the first time, that a novel plant-derived nanofibrillar cellulose (NFC) hydrogel creates a flexible 3D environment for hPSC culture. The pluripotency of hPSCs cultured in the NFC hydrogel was maintained for 26 days as evidenced by the expression of OCT4, NANOG, and SSEA-4, in vitro embryoid body formation and in vivo teratoma formation. The use of a cellulose enzyme, cellulase, enables easy cell propagation in 3D culture as well as a shift between 3D and two-dimensional cultures. More importantly, the removal of the NFC hydrogel facilitates differentiation while retaining 3D cell organization. Thus, the NFC hydrogel represents a flexible, xeno-free 3D culture system that supports pluripotency and will be useful in hPSC-based drug research and regenerative medicine.
authors.affiliation: 1 Division of Biopharmaceutics and Pharmacokinetics, Faculty of Pharmacy, University of Helsinki , Helsinki, Finland .
authors.forename: Yan-Ru
authors.initials: YR
authors.lastname: Lou
authors.forename: Liisa
authors.initials: L
authors.lastname: Kanninen
authors.forename: Tytti
authors.initials: T
authors.lastname: Kuisma
authors.forename: Johanna
authors.initials: J
authors.lastname: Niklander
authors.forename: Luke A
authors.initials: LA
authors.lastname: Noon
authors.forename: Deborah
authors.initials: D
authors.lastname: Burks
authors.forename: Arto
authors.initials: A
authors.lastname: Urtti
authors.forename: Marjo
authors.initials: M
authors.lastname: Yliperttula
mesh_terms: D018929:Cell Culture Techniques; D002454:Cell Differentiation; D002460:Cell Line; D002482:Cellulose / Q000737:chemistry*; D003470:Culture Media / Q000737:chemistry; D018398:Homeodomain Proteins / Q000378:metabolism; D006801:Humans; D020100:Hydrogels / Q000737:chemistry*; D057026:Induced Pluripotent Stem Cells / Q000502:physiology*; D059785:Karyotype; D008928:Mitochondria / Q000378:metabolism; D057139:Nanofibers / Q000737:chemistry*; D000071317:Nanog Homeobox Protein; D050814:Octamer Transcription Factor-3 / Q000378:metabolism; D018874:Spheroids, Cellular / Q000502:physiology
chemical_list: D003470:Culture Media; D018398:Homeodomain Proteins; D020100:Hydrogels; C486780:NANOG protein, human; D000071317:Nanog Homeobox Protein; D050814:Octamer Transcription Factor-3; C494173:POU5F1 protein, human; D002482:Cellulose