Control of neural cell composition in poly(ethylene glycol) hydrogel culture with soluble factors.

Poly(ethylene glycol) (PEG) hydrogels are being developed as cell delivery vehicles that have great potential to improve neuronal replacement therapies. Current research priorities include (1) characterizing neural cell growth within PEG hydrogels relative to standard culture systems and (2) generating neuronal-enriched populations within the PEG hydrogel environment. This study compares the percentage of neural precursor cells (NPCs), neurons, and glia present when dissociated neural cells are seeded within PEG hydrogels relative to standard monolayer culture. Results demonstrate that PEG hydrogels enriched the initial cell population for NPCs, which subsequently gave rise to neurons, then to glia. Relative to monolayer culture, PEG hydrogels maintained an increased percentage of NPCs and a decreased percentage of glia. This neurogenic advantage of PEG hydrogels is accentuated in the presence of basic fibroblast growth factor and epidermal growth factor, which more potently increase NPC and neuronal expression markers when applied to cells cultured within PEG hydrogels. Finally, this work demonstrates that glial differentiation can be selectively eliminated upon supplementation with a γ-secretase inhibitor. Together, this study furthers our understanding of how the PEG hydrogel environment influences neural cell composition and also describes select soluble factors that are useful in generating neuronal-enriched populations within the PEG hydrogel environment.

[1]  Kristi S Anseth,et al.  Effects of PEG hydrogel crosslinking density on protein diffusion and encapsulated islet survival and function. , 2009, Journal of biomedical materials research. Part A.

[2]  David F Meaney,et al.  Matrices with compliance comparable to that of brain tissue select neuronal over glial growth in mixed cortical cultures. , 2006, Biophysical journal.

[3]  C. Santoro,et al.  Activation of caspase-8 triggers anoikis in human neuroblastoma cells , 2006, Neuroscience Research.

[4]  Andrés J. García,et al.  Specific β1 integrins mediate adhesion, migration, and differentiation of neural progenitors derived from the embryonic striatum , 2004, Molecular and Cellular Neuroscience.

[5]  E. Lavik,et al.  Fabrication of degradable polymer scaffolds to direct the integration and differentiation of retinal progenitors. , 2005, Biomaterials.

[6]  M. Mahoney,et al.  Biocompatibility of PEG-Based Hydrogels in Primate Brain , 2008, Cell transplantation.

[7]  A. Cressant,et al.  Homotopic septal grafts combined with a hydrogel bridge promote functional recovery in rats with fimbria-fornix lesions: A unit recording study. , 1999, Restorative neurology and neuroscience.

[8]  L. Romer,et al.  Integrin-Mediated Survival Signals Regulate the Apoptotic Function of Bax through Its Conformation and Subcellular Localization , 2000, The Journal of cell biology.

[9]  S. Bryant,et al.  Hydrogel properties influence ECM production by chondrocytes photoencapsulated in poly(ethylene glycol) hydrogels. , 2002, Journal of biomedical materials research.

[10]  F. Guillemot,et al.  Transcription factors Mash-1 and Prox-1 delineate early steps in differentiation of neural stem cells in the developing central nervous system. , 1999, Development.

[11]  M. Mahoney,et al.  Impact of lactic acid on cell proliferation and free radical‐induced cell death in monolayer cultures of neural precursor cells , 2009, Biotechnology and bioengineering.

[12]  D. Kooy,et al.  Hematopoietic competence is a rare property of neural stem cells that may depend on genetic and epigenetic alterations , 2002, Nature Medicine.

[13]  N. Quarto,et al.  Heparan sulfate proteoglycans as transducers of FGF-2 signalling. , 1994, Journal of cell science.

[14]  R. Cassens,et al.  Comparison of Ribogreen ® and 18s Rrna Quantitation for Normalizing Real-time Rt-pcr Expression Analysis , 2022 .

[15]  M. Mahoney,et al.  Development of porous PEG hydrogels that enable efficient, uniform cell-seeding and permit early neural process extension. , 2009, Acta biomaterialia.

[16]  A S Hoffman,et al.  Reduction of fibrinogen adsorption on PEG-coated polystyrene surfaces. , 1992, Journal of biomedical materials research.

[17]  J G Parnavelas,et al.  Collagen type IV promotes the differentiation of neuronal progenitors and inhibits astroglial differentiation in cortical cell cultures. , 1998, Brain research. Developmental brain research.

[18]  Olle Inganäs,et al.  The promotion of neuronal maturation on soft substrates. , 2009, Biomaterials.

[19]  S. Morrison,et al.  Physiological Notch signaling promotes gliogenesis in the developing peripheral and central nervous systems , 2007, Development.

[20]  T. Shimazaki,et al.  Prospective characterization of neural stem cells by flow cytometry analysis using a combination of surface markers , 2005, Journal of neuroscience research.

[21]  Jeffrey A. Hubbell,et al.  Bioerodible hydrogels based on photopolymerized poly(ethylene glycol)-co-poly(.alpha.-hydroxy acid) diacrylate macromers , 1993 .

[22]  K. Heidenreich,et al.  IGF-I and bFGF Improve Dopamine Neuron Survival and Behavioral Outcome in Parkinsonian Rats Receiving Cultured Human Fetal Tissue Strands , 2001, Experimental Neurology.

[23]  A. Björklund,et al.  Cell replacement therapies for central nervous system disorders , 2000, Nature Neuroscience.

[24]  Tong Zheng,et al.  Production and analysis of neurospheres from acutely dissociated and postmortem CNS specimens. , 2002, Methods in molecular biology.

[25]  S. Jo,et al.  Presenilin 1/gamma-secretase is associated with cadmium-induced E-cadherin cleavage and COX-2 gene expression in T47D breast cancer cells. , 2008, Toxicological sciences : an official journal of the Society of Toxicology.

[26]  R E Baier,et al.  Conditioning surfaces to suit the biomedical environment: recent progress. , 1982, Journal of biomechanical engineering.

[27]  A Streptomyces fradiae protease dissociates structurally preserved neurons and glial cells from the embryonic and adult central nervous system of vertebrates , 1997, Journal of Neuroscience Methods.

[28]  M. Dickinson,et al.  Quiescence and Activation of Stem and Precursor Cell Populations in the Subependymal Zone of the Mammalian Brain Are Associated with Distinct Cellular and Extracellular Matrix Signals , 2010, The Journal of Neuroscience.

[29]  U. Rodeck,et al.  Matrix-independent survival of human keratinocytes through an EGF receptor/MAPK-kinase-dependent pathway. , 2001, Molecular biology of the cell.

[30]  S. Temple Division and differentiation of isolated CNS blast cells in microculture , 1989, Nature.

[31]  K. Anseth,et al.  The effect of heparin-functionalized PEG hydrogels on three-dimensional human mesenchymal stem cell osteogenic differentiation. , 2007, Biomaterials.

[32]  A. Czurkó,et al.  Reconstruction of GABAergic Transmission and Behavior by Striatal Cell Grafts in Rats with Ischemic Infarcts in the Middle Cerebral Artery , 1993, Journal of neural transplantation & plasticity.

[33]  S. Goderie,et al.  Timing of CNS Cell Generation A Programmed Sequence of Neuron and Glial Cell Production from Isolated Murine Cortical Stem Cells , 2000, Neuron.

[34]  F. Gage,et al.  Mammalian neural stem cells. , 2000, Science.

[35]  C. Werner,et al.  Analytical approaches to uptake and release of hydrogel-associated FGF-2 , 2010, Journal of materials science. Materials in medicine.

[36]  B. Matthews,et al.  Three-dimensional structure of human basic fibroblast growth factor. , 1991, Proceedings of the National Academy of Sciences of the United States of America.

[37]  Matthias P Lutolf,et al.  Enhancing the Reliability and Throughput of Neurosphere Culture on Hydrogel Microwell Arrays , 2008, Stem cells.

[38]  C. Streuli,et al.  Early Events in the Anoikis Program Occur in the Absence of Caspase Activation* , 2003, Journal of Biological Chemistry.

[39]  D. van der Kooy,et al.  Notch pathway molecules are essential for the maintenance, but not the generation, of mammalian neural stem cells. , 2002, Genes & development.

[40]  G. Buettner The pecking order of free radicals and antioxidants: lipid peroxidation, alpha-tocopherol, and ascorbate. , 1993, Archives of biochemistry and biophysics.

[41]  M. Mahoney,et al.  Impact of degradable macromer content in a poly(ethylene glycol) hydrogel on neural cell metabolic activity, redox state, proliferation, and differentiation. , 2010, Tissue engineering. Part A.

[42]  A. Ghosh,et al.  Sequential specification of neurons and glia by developmentally regulated extracellular factors. , 2001, Development.

[43]  A. Björklund,et al.  Intracerebral grafting of dissociated CNS tissue suspensions: a new approach for neuronal transplantation to deep brain sites , 1981, Brain Research.

[44]  D. van der Kooy,et al.  Distinct neural stem cells proliferate in response to EGF and FGF in the developing mouse telencephalon. , 1999, Developmental biology.

[45]  Kyle J. Lampe,et al.  Effect of macromer weight percent on neural cell growth in 2D and 3D nondegradable PEG hydrogel culture. , 2010, Journal of biomedical materials research. Part A.

[46]  M. Filla,et al.  The cell surface proteoglycan syndecan‐1 mediates fibroblast growth factor‐2 binding and activity , 1998, Journal of cellular physiology.

[47]  Lia S. Campos,et al.  β1 integrins activate a MAPK signalling pathway in neural stem cells that contributes to their maintenance , 2004, Development.

[48]  T. Golde,et al.  γ-Secretase Cleavage and Nuclear Localization of ErbB-4 Receptor Tyrosine Kinase , 2001, Science.

[49]  M. Mahoney,et al.  Specific fibrinogen and thrombin concentrations promote neuronal rather than glial growth when primary neural cells are seeded within plasma-derived fibrin gels. , 2010, Tissue engineering. Part A.

[50]  W. Saltzman,et al.  Cultures of cells from fetal rat brain: methods to control composition, morphology, and biochemical activity. , 1999, Biotechnology and bioengineering.

[51]  Erin B Lavik,et al.  Photopolymerized poly(ethylene glycol)/poly(L-lysine) hydrogels for the delivery of neural progenitor cells , 2007, Journal of biomaterials science. Polymer edition.

[52]  Kristi S Anseth,et al.  Three-dimensional growth and function of neural tissue in degradable polyethylene glycol hydrogels. , 2006, Biomaterials.

[53]  F. Gage,et al.  Functional neuronal replacement by grafted striatal neurones in the ibotenic acid-lesioned rat striatum , 1984, Nature.

[54]  C. Haass,et al.  A γ‐secretase inhibitor blocks Notch signaling in vivo and causes a severe neurogenic phenotype in zebrafish , 2002, EMBO reports.

[55]  D. Seliktar,et al.  Biosynthetic hydrogel scaffolds made from fibrinogen and polyethylene glycol for 3D cell cultures. , 2005, Biomaterials.

[56]  A. Hampl,et al.  Inhibition of Notch Signaling in Human Embryonic Stem Cell–Derived Neural Stem Cells Delays G1/S Phase Transition and Accelerates Neuronal Differentiation In Vitro and In Vivo , 2010, Stem cells.

[57]  F. Cardozo-Pelaez,et al.  Comparison of base-excision repair capacity in proliferating and differentiated PC 12 cells following acute challenge with dieldrin. , 2001, Free radical biology & medicine.

[58]  K. Anseth,et al.  Contrasting effects of collagen and bFGF-2 on neural cell function in degradable synthetic PEG hydrogels. , 2007, Journal of biomedical materials research. Part A.

[59]  K. Nakanishi,et al.  Identification and Functions of Chondroitin Sulfate in the Milieu of Neural Stem Cells* , 2006, Journal of Biological Chemistry.

[60]  I. Ferguson,et al.  Basic fibroblast growth factor: receptor-mediated internalization, metabolism, and anterograde axonal transport in retinal ganglion cells , 1990, The Journal of neuroscience : the official journal of the Society for Neuroscience.

[61]  Fengqin Gao,et al.  Bcl2 retards G1/S cell cycle transition by regulating intracellular ROS. , 2003, Blood.

[62]  H. Roelink,et al.  The Notch response inhibitor DAPT enhances neuronal differentiation in embryonic stem cell‐derived embryoid bodies independently of sonic hedgehog signaling , 2007, Developmental dynamics : an official publication of the American Association of Anatomists.

[63]  Austin G Smith,et al.  Niche-Independent Symmetrical Self-Renewal of a Mammalian Tissue Stem Cell , 2005, PLoS biology.

[64]  James W. Fawcett,et al.  Building a Bridge: Engineering Spinal Cord Repair , 2002, Experimental Neurology.