Neural stem/progenitor cells cultured on gamma-crosslinked poly(vinyl alcohol) hydrogels

Abstract

Event Abstract Back to Event Neural stem/progenitor cells cultured on gamma-crosslinked poly(vinyl alcohol) hydrogels Hideki Mori1* and Masayuki Hara1* 1 Osaka Prefecture Univertisy, Department of Biological Science, Graduate School of Science,, Japan Introduction: Neural stem/progenitor cells (NSPCs) in the central nervous system (CNS) have the capacity to self-renew by proliferation and are multipotent, giving rise to neurons, astrocytes, and oligodendrocytes. NSPCs can be amplified in neurosphere suspension cultures for cell transplantation therapy to treat CNS diseases as well as for in vitro pharmacological/toxicological assays; however, these suspension cultures have certain limitations, including the inconvenience of changing the culture medium as well as difficulty of live imaging. In the present study, we prepared a gamma-crosslinked poly(vinyl alcohol) (PVA) hydrogel and assessed its suitability as a substrate for adherent NSPC cultures. Experimental: An aqueous solution of PVA at concentrations of 3.75% (w/v), 7.50 % (w/v), or 15.0 % (w/v) was irradiated with various doses (10, 20, or 40 kGy) of gamma ray at room temperature (25°C) in the cobalt 60 (60Co) gamma ray facility of the Radiation Research Center of Osaka Prefecture University. NSPCs were isolated from the fetal brain of an imprinting control region mouse (embryonic day 14) and resuspended in DMEM/F-12 medium supplemented with EGF (20 ng/ml), b-FGF (20 ng/ml), and B-27 supplement. Cells (1.0 × 105/ml in 10 ml medium) were cultured in dishes at 37°C and 5% (v/v) CO2 to induce neurosphere formation and were expanded by dissociation with trypsin and reseeding every 7 days. All animal experiments were conducted in accordance with institutional guidelines and national standards with approval from the Animal Experiment Committee of Osaka Prefecture University. NSPCs amplified for several passages as described above were used for analysis on a PVA gel. Differentiation was determined by evaluating the expression of the markers nestin (progenitors), βIII tubulin (neurons), and glial fibrillary acidic protein and S100 β (glia) by immunocytochemistry and quantitative reverse transcriptase PCR. Results and Discussion: NSPCs adhered to the PVA gel as clusters and grew without differentiating into neurons and glia. The cluster was hemisphere or in a mount-like shape on the surface of the PVA gel. The proliferation rate of cells grown on the soft PVA gel (3.75-7.5% (w/v) PVA) was approximately 70% of that of neurospheres in suspension. The levels of the marker genes were similar between the two types of culture; although some variability was observed, there were no fold differences in expression. We conclude that gamma-crosslinked PVA hydrogels can function as a novel scaffold for maintaining adherent NSPCs in an undifferentiated state. Keywords: Cell Adhesion, Cell Differentiation, stem cell, Biocompatibility Conference: 10th World Biomaterials Congress, Montréal, Canada, 17 May - 22 May, 2016. Presentation Type: Poster Topic: Regenerative medicine: biomaterials for control of tissue induction Citation: Mori H and Hara M (2016). Neural stem/progenitor cells cultured on gamma-crosslinked poly(vinyl alcohol) hydrogels. Front. Bioeng. Biotechnol. Conference Abstract: 10th World Biomaterials Congress. doi: 10.3389/conf.FBIOE.2016.01.01540 Copyright: The abstracts in this collection have not been subject to any Frontiers peer review or checks, and are not endorsed by Frontiers. They are made available through the Frontiers publishing platform as a service to conference organizers and presenters. The copyright in the individual abstracts is owned by the author of each abstract or his/her employer unless otherwise stated. Each abstract, as well as the collection of abstracts, are published under a Creative Commons CC-BY 4.0 (attribution) licence (https://creativecommons.org/licenses/by/4.0/) and may thus be reproduced, translated, adapted and be the subject of derivative works provided the authors and Frontiers are attributed. For Frontiers’ terms and conditions please see https://www.frontiersin.org/legal/terms-and-conditions. Received: 27 Mar 2016; Published Online: 30 Mar 2016. * Correspondence: Dr. Hideki Mori, Osaka Prefecture Univertisy, Department of Biological Science, Graduate School of Science,, Sakai-city, Osaka Prefecture, Japan, morihide@b.s.osakafu-u.ac.jp Dr. Masayuki Hara, Osaka Prefecture Univertisy, Department of Biological Science, Graduate School of Science,, Sakai-city, Osaka Prefecture, Japan, hara@b.s.osakafu-u.ac.jp Login Required This action requires you to be registered with Frontiers and logged in. To register or login click here. 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