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Optimization of fibrin scaffolds for differentiation of murine embryonic stem cells into neural lineage cells

Paper ID Volume ID Publish Year Pages File Format Full-Text
11268 729 2006 14 PDF Available
Title
Optimization of fibrin scaffolds for differentiation of murine embryonic stem cells into neural lineage cells
Abstract

The 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 and astrocytes 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.

Keywords
Nerve tissue engineering; Cell spreading; Cell culture; Progenitor cells
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Optimization of fibrin scaffolds for differentiation of murine embryonic stem cells into neural lineage cells
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Publisher
Database: Elsevier - ScienceDirect
Journal: Biomaterials - Volume 27, Issue 36, December 2006, Pages 5990–6003
Authors
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Subjects
Physical Sciences and Engineering Chemical Engineering Bioengineering
Get Full-Text Now
Don't Miss Today's Special Offer
Price was $35.95
You save - $31
Price after discount Only $4.95
100% Money Back Guarantee
Full-text PDF Download
Online Support
Any Questions? feel free to contact us