title: Gelatin-PMVE/MA composite scaffold promotes expansion of embryonic stem cells.
abstract: We introduce a new composite scaffold of gelatin and polymethyl vinyl ether-alt-maleic anhydride (PMVE/MA) for expansion of embryonic stem cells (ESCs) in an in vitro environment. To optimize the scaffold, we prepared a gelatin scaffold (G) and three composite scaffolds namely GP-1, GP-2, and GP-3 with varying PMVE/MA concentrations (0.2-1%) and characterized them by scanning electron microscopy (SEM), swelling study, compression testing and FTIR. SEM micrographs revealed interconnected porous structure in all the scaffolds. The permissible hemolysis ratio and activation of platelets by scaffolds confirmed the hemocompatibility of scaffolds. Initial biocompatibility assessment of scaffolds was conducted using hepatocarcinoma (Hep G2) cells and adhesion, proliferation and infiltration of Hep G2 cells in depth of scaffolds were observed, proving the scaffold's biocompatibility. Further Oct4B2 mouse embryonic stem cells (mESCs), which harbor a green fluorescence protein transgene under regulatory control of the Oct4 promotor, were examined for expansion on scaffolds with MTT assay. The GP-2 scaffold demonstrated the best cell proliferation and was further explored for ESC adherence and infiltration in depth (SEM and confocal), and pluripotent state of mESCs was assessed with the expression of Oct4-GFP and stage-specific embryonic antigen-1 (SSEA-1). This study reports the first demonstration of biocompatibility of gelatin-PMVE/MA composite scaffold and presents this scaffold as a promising candidate for embryonic stem cell based tissue engineering.
authors.affiliation: Department of Chemical Engineering, Indian Institute of Technology Bombay, Powai, Mumbai, India.
authors.forename: Hemlata
authors.initials: H
authors.lastname: Chhabra
authors.affiliation: Department of Chemical Engineering, Indian Institute of Technology Bombay, Powai, Mumbai, India; IITB-Monash Research Academy, Mumbai, India; Department of Chemical Engineering, Monash University, Melbourne, Australia.
authors.forename: Priyanka
authors.initials: P
authors.lastname: Gupta
authors.affiliation: Turretfield Research Centre, South Australian Research and Development Institute, Rosedale, South Australia, Australia.
authors.forename: Paul J
authors.initials: PJ
authors.lastname: Verma
authors.affiliation: Department of Chemical Engineering, Indian Institute of Technology Bombay, Powai, Mumbai, India.
authors.forename: Sameer
authors.initials: S
authors.lastname: Jadhav
authors.affiliation: Department of Chemical Engineering, Indian Institute of Technology Bombay, Powai, Mumbai, India.
authors.forename: Jayesh R
authors.initials: JR
authors.lastname: Bellare
mesh_terms: D000818:Animals; D001672:Biocompatible Materials / Q000737:chemistry* / Q000494:pharmacology; D049109:Cell Proliferation / Q000187:drug effects; D002478:Cells, Cultured; D053595:Embryonic Stem Cells / Q000166:cytology* / Q000378:metabolism; D005780:Gelatin / Q000737:chemistry*; D056945:Hep G2 Cells; D006801:Humans; D016256:Lewis X Antigen / Q000378:metabolism; D008298:Maleates / Q000737:chemistry*; D051379:Mice; D050814:Octamer Transcription Factor-3 / Q000378:metabolism; D011095:Polyethylenes / Q000737:chemistry*; D016062:Porosity; D054457:Tissue Scaffolds
chemical_list: D001672:Biocompatible Materials; D016256:Lewis X Antigen; D008298:Maleates; D050814:Octamer Transcription Factor-3; D011095:Polyethylenes; C494174:Pou5f1 protein, mouse; D005780:Gelatin; C043105:poly(methyl vinyl ether-co-maleic anhydride)
keywords: Biocompatibility; Cell infiltration; Embryonic stem cell; Gelatin; Pluripotency; Polymethyl vinyl ether-alt-maleic anhydride