title: Expansion and differentiation of germline-derived pluripotent stem cells on biomaterials.
abstract: Stem cells with broad differentiation potential, such as the recently described germline-derived pluripotent stem cells (gPS cells), are an appealing source for tissue engineering strategies. Biomaterials can inhibit, support, or induce proliferation and differentiation of stem cells. Here we identified (1) polymers that maintain self-renewal and differentiation potential of gPS cells for feeder-free expansion and (2) polymers supporting the cardiomyogenic fate of gPS cells by analyzing a panel of polymers of an established biomaterial bank previously used to assess growth of diverse stem cell types. Identification of cytocompatible gPS cell/biomaterial combinations required analysis of several parameters, including morphology, viability, cytotoxicity, apoptosis, proliferation, and differentiation potential. Pluripotency of gPS cells was visualized by the endogenous Oct4-promoter-driven GFP and by Sox2 and Nanog immunofluorescence. Viability assay, proliferation assay, and flow cytometry showed that gPS cells efficiently adhere and are viable on synthetic polymers, such as Resomer(®) LR704 (poly(L-lactic-D,L-lactic acid), poly(tetrafluor ethylene) (PTFE), poly(vinylidene fluoride) (PVDF), and on gelatine-coated tissue culture polystyrene. Expansion experiments showed that Resomer LR704 is an alternative substrate for feeder-free gPS cell maintenance. Resomer LR704, PTFE, and PVDF were found to be suitable for gPS cell differentiation. Spontaneous beating in embryoid bodies cultured on Resomer LR704 occurred already on day 8 of differentiation, much earlier compared to the other surfaces. This indicates that Resomer LR704 supports spontaneous cardiomyogenic differentiation of gPS cells, which was also confirmed on molecular, protein and functional level.
authors.affiliation: Interdisciplinary Centre for Clinical Research Aachen IZKF Aachen, RWTH Aachen University, Aachen 52074, Germany. mareike.hoss@rwth-aachen.de
authors.forename: Mareike
authors.initials: M
authors.lastname: Hoss
authors.forename: Tomo
authors.initials: T
authors.lastname: Šarić
authors.forename: Bernd
authors.initials: B
authors.lastname: Denecke
authors.forename: Gabriel
authors.initials: G
authors.lastname: Peinkofer
authors.forename: Manfred
authors.initials: M
authors.lastname: Bovi
authors.forename: Jürgen
authors.initials: J
authors.lastname: Groll
authors.forename: Kinarm
authors.initials: K
authors.lastname: Ko
authors.forename: Jochen
authors.initials: J
authors.lastname: Salber
authors.forename: Marcel
authors.initials: M
authors.lastname: Halbach
authors.forename: Hans R
authors.initials: HR
authors.lastname: Schöler
authors.forename: Martin
authors.initials: M
authors.lastname: Zenke
authors.forename: Sabine
authors.initials: S
authors.lastname: Neuss
mesh_terms: D000818:Animals; D001672:Biocompatible Materials / Q000737:chemistry*; D002454:Cell Differentiation / Q000502:physiology; D049109:Cell Proliferation; D002478:Cells, Cultured; D005434:Flow Cytometry; D005854:Germ Cells; D007150:Immunohistochemistry; D019344:Lactic Acid / Q000737:chemistry; D051379:Mice; D008855:Microscopy, Electron, Scanning; D039904:Pluripotent Stem Cells / Q000166:cytology* / Q000378:metabolism; D011091:Polyesters; D011108:Polymers / Q000737:chemistry; D011138:Polytetrafluoroethylene / Q000737:chemistry; D011145:Polyvinyls / Q000737:chemistry; D020133:Reverse Transcriptase Polymerase Chain Reaction; D005465:Fluorocarbon Polymers
chemical_list: D001672:Biocompatible Materials; D019344:Lactic Acid; D011091:Polyesters; D011108:Polymers; D011138:Polytetrafluoroethylene; D011145:Polyvinyls; C024865:polyvinylidene fluoride; D005465:Fluorocarbon Polymers; C033616:poly(lactide)