Optimization of Fibrin Scaffolds for Differentiation of Murine Embryonic Stem Cells into Neural Lineage Cells

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dc.contributor.authorWillerth, Stephanie
dc.contributor.authorArendas, Kelly J
dc.contributor.authorGottlieb, David I
dc.contributor.authorSakiyama-Elbert, Shelly E
dc.date.accessioned2015-07-03T16:59:52Z
dc.date.available2015-07-03T16:59:52Z
dc.date.copyright2006en_US
dc.date.issued2006-12
dc.descriptionAuthor manuscript, post peer review (post-print)en_US
dc.description.abstractThe objective of this research was to determine the appropriate cell culture conditions for embryonic stem (ES) cell proliferation and differentiation in fibrin scaffolds by examining cell seeding density, location, and the optimal concentrations of fibrinogen, thrombin, and aprotinin (protease inhibitor). Mouse ES cells were induced to become neural progenitors by adding retinoic acid for 4 days to embryoid body (EB) cultures. For dissociated EBs, the optimal cell seeding density and location was determined to be 250,000 cells/cm² seeded on top of fibrin scaffolds. For intact EBs, three dimensional (3D) cultures with one EB per 400 μL fibrin scaffold resulted in greater cell proliferation and differentiation than two dimensional (2D) cultures. Optimal concentrations for scaffold polymerization were 10 mg/mL of fibrinogen and 2 NIH units/mL of thrombin. The optimal aprotinin concentration was determined to be 50 μg/mL for dissociated EBs (2D) and 5 μg/mL for intact EBs in 3D fibrin scaffolds. Additionally, after 14 days in 3D culture EBs differentiated into neurons andastrocytes as indicated by immunohistochemisty. These conditions provide an optimal fibrin scaffold for evaluating ES cell differentiation and proliferation in culture, and for use as a platform for neural tissue engineering applications, such as the treatment for spinal cord injury.en_US
dc.description.reviewstatusRevieweden_US
dc.description.scholarlevelFacultyen_US
dc.description.sponsorshipThe authors acknowledge NIH grant NS051454, which provided the funding for this work.en_US
dc.identifier.citationWillerth, S.M. et al (2006) Optimization of Fibrin Scaffolds for Differentiation of Murine Embryonic Stem Cells into Neural Lineage Cells. Biomaterials 27(36): 5990–6003.en_US
dc.identifier.urihttp://dx.doi.org/10.1016/j.biomaterials.2006.07.036
dc.identifier.urihttp://hdl.handle.net/1828/6286
dc.language.isoenen_US
dc.publisherElsevieren_US
dc.rightsAttribution-NonCommercial-NoDerivs 2.5 Canada*
dc.rights.urihttp://creativecommons.org/licenses/by-nc-nd/2.5/ca/*
dc.subjectnerve tissue engineering
dc.subjectcell spreading
dc.subjectcell culture
dc.subjectprogenitor cells
dc.subject.departmentDepartment of Mechanical Engineering
dc.subject.departmentSchool of Medical Sciences
dc.titleOptimization of Fibrin Scaffolds for Differentiation of Murine Embryonic Stem Cells into Neural Lineage Cellsen_US
dc.typePreprinten_US

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