title: Nanotopographical control of human embryonic stem cell differentiation into definitive endoderm.
abstract: Derivation of definitive endoderm (DE) from human embryonic stem cells (hESCs) can address the needs of regenerative medicine for endoderm-derived organs such as the pancreas and liver. Fibrous substrates which topographically recapitulate native extracellular matrix have been known to promote the stem cell differentiation. However, the optimal fiber diameter remains to be determined for the desired differentiation. Here, we have developed a simple method to precisely fabricate electrospun poly(ε-caprolactone) fibers with four distinct average diameters at nano- and microscale levels (200, 500, 800, and 1300 nm). Human ESCs were cultured as clumps or single cells and induced into DE differentiation to determine the optimal topography leading to the promoted differentiation compared with planar culture plates. Gene expression analysis of the DE-induced cells showed significant upregulation of DE-specific genes exclusively on the 200-nm fibers. By Western blot analysis, significant expression of DE-specific proteins was found when hESCs were cultured on the 200 nm substrate as single cells rather than clumps, probably due to more efficient cell-matrix interaction realized by morphological observations of the cell colonies. The results indicated that nanofibrillar substrates, only at ultrathin fiber diameters, provided a better environment for DE differentiation of hESC, which holds great promise in prospective tissue engineering applications.
authors.affiliation: Department of Stem Cells and Developmental Biology at Cell Science Research Center, Royan Institute for Stem Cell Biology and Technology, ACECR, Tehran, Iran.|Biomaterials Department, Iran Polymer and Petrochemical Institute, Tehran, Iran.
authors.forename: Mohammad Hossein
authors.initials: MH
authors.lastname: Ghanian
authors.affiliation: Department of Stem Cells and Developmental Biology at Cell Science Research Center, Royan Institute for Stem Cell Biology and Technology, ACECR, Tehran, Iran.
authors.forename: Zahra
authors.initials: Z
authors.lastname: Farzaneh
authors.affiliation: Biomaterials Department, Iran Polymer and Petrochemical Institute, Tehran, Iran.
authors.forename: Jalal
authors.initials: J
authors.lastname: Barzin
authors.affiliation: Biomaterials Department, Iran Polymer and Petrochemical Institute, Tehran, Iran.
authors.forename: Mojgan
authors.initials: M
authors.lastname: Zandi
authors.affiliation: Department of Stem Cells and Developmental Biology at Cell Science Research Center, Royan Institute for Stem Cell Biology and Technology, ACECR, Tehran, Iran.|Biomaterials Department, Iran Polymer and Petrochemical Institute, Tehran, Iran.
authors.forename: Mohammad
authors.initials: M
authors.lastname: Kazemi-Ashtiani
authors.affiliation: Department of Stem Cells and Developmental Biology at Cell Science Research Center, Royan Institute for Stem Cell Biology and Technology, ACECR, Tehran, Iran.
authors.forename: Mehdi
authors.initials: M
authors.lastname: Alikhani
authors.affiliation: Biomaterials Department, Iran Polymer and Petrochemical Institute, Tehran, Iran.
authors.forename: Morteza
authors.initials: M
authors.lastname: Ehsani
authors.affiliation: Department of Stem Cells and Developmental Biology at Cell Science Research Center, Royan Institute for Stem Cell Biology and Technology, ACECR, Tehran, Iran.|Department of Developmental Biology, University of Science and Culture, ACECR, Tehran, Iran.
authors.forename: Hossein
authors.initials: H
authors.lastname: Baharvand
mesh_terms: D002449:Cell Aggregation / Q000187:drug effects; D002454:Cell Differentiation / Q000187:drug effects; D048430:Cell Shape / Q000187:drug effects; D003114:Colony-Forming Units Assay; D004707:Endoderm / Q000166:cytology*; D000066449:Human Embryonic Stem Cells / Q000166:cytology*; D006801:Humans; D053758:Nanoparticles / Q000737:chemistry* / Q000648:ultrastructure; D011091:Polyesters / Q000494:pharmacology; D016062:Porosity; D060888:Real-Time Polymerase Chain Reaction; D012995:Solubility; D012997:Solvents
chemical_list: D011091:Polyesters; D012997:Solvents; C016240:polycaprolactone
keywords: definitive endoderm; differentiation; electrospinning; human embryonic stem cell; topography