title: A synthetic, xeno-free peptide surface for expansion and directed differentiation of human induced pluripotent stem cells.
abstract: Human induced pluripotent stem cells have the potential to become an unlimited cell source for cell replacement therapy. The realization of this potential, however, depends on the availability of culture methods that are robust, scalable, and use chemically defined materials. Despite significant advances in hiPSC technologies, the expansion of hiPSCs relies upon the use of animal-derived extracellular matrix extracts, such as Matrigel, which raises safety concerns over the use of these products. In this work, we investigated the feasibility of expanding and differentiating hiPSCs on a chemically defined, xeno-free synthetic peptide substrate, i.e. Corning Synthemax(®) Surface. We demonstrated that the Synthemax Surface supports the attachment, spreading, and proliferation of hiPSCs, as well as hiPSCs' lineage-specific differentiation. hiPSCs colonies grown on Synthemax Surfaces exhibit less spread and more compact morphology compared to cells grown on Matrigel™. The cytoskeleton characterization of hiPSCs grown on the Synthemax Surface revealed formation of denser actin filaments in the cell-cell interface. The down-regulation of vinculin and up-regulation of zyxin expression were also observed in hiPSCs grown on the Synthemax Surface. Further examination of cell-ECM interaction revealed that hiPSCs grown on the Synthemax Surface primarily utilize α(v)β(5) integrins to mediate attachment to the substrate, whereas multiple integrins are involved in cell attachment to Matrigel. Finally, hiPSCs can be maintained undifferentiated on the Synthemax Surface for more than ten passages. These studies provide a novel approach for expansion of hiPSCs using synthetic peptide engineered surface as a substrate to avoid a potential risk of contamination and lot-to-lot variability with animal derived materials.
authors.affiliation: Department of Biomedical Engineering, College of Engineering, University of Arkansas, Fayetteville, AR, USA. sjin@uark.edu
authors.forename: Sha
authors.initials: S
authors.lastname: Jin
authors.forename: Huantong
authors.initials: H
authors.lastname: Yao
authors.forename: Jennifer L
authors.initials: JL
authors.lastname: Weber
authors.forename: Zara K
authors.initials: ZK
authors.lastname: Melkoumian
authors.forename: Kaiming
authors.initials: K
authors.lastname: Ye
mesh_terms: D002448:Cell Adhesion / Q000187:drug effects; D002454:Cell Differentiation / Q000187:drug effects*; D049109:Cell Proliferation / Q000187:drug effects; D003094:Collagen / Q000494:pharmacology; D003599:Cytoskeleton / Q000187:drug effects / Q000378:metabolism; D004338:Drug Combinations; D004707:Endoderm / Q000166:cytology; D006801:Humans; D057026:Induced Pluripotent Stem Cells / Q000166:cytology* / Q000187:drug effects* / Q000378:metabolism; D007797:Laminin / Q000494:pharmacology; D010455:Peptides / Q000494:pharmacology*; D011509:Proteoglycans / Q000494:pharmacology; D019030:Receptors, Vitronectin / Q000378:metabolism
chemical_list: D004338:Drug Combinations; D007797:Laminin; D010455:Peptides; D011509:Proteoglycans; D019030:Receptors, Vitronectin; C064660:integrin alphaVbeta5; C059485:matrigel; D003094:Collagen