Three-dimensional Culture for Expansion and Differentiation of Mouse Embryonic Stem Cells.
From: Zimmer Inc., 12024 Vista Parke Drive, Austin, TX 78726, USA.
Biomaterials
- Publish Date: Dec 2006
- ISSN: 0142-9612
- Volume: 27
- Issue: 36
- Pages: 6004-14
- Medium: Print
- Language: English
- Citation (JAMA): Liu Hui, Collins Scott F, Suggs Laura J, et al. Three-dimensional Culture for Expansion and Differentiation of Mouse Embryonic Stem Cells.. Biomaterials Dec 2006;27:6004-14
Abstract
Differentiation of embryonic stem (ES) cells typically requires cell-cell aggregation in the form of embryoid bodies (EBs). This process is not very well controlled and final cell numbers can be limited by EB agglomeration and the inability to drive differentiation towards a desired cell type. This study compares three-dimensional (3D) fibrin culture to conventional two-dimensional (2D) suspension culture and to culture in a semisolid methylcellulose medium solution. Two types of fibrin culture were evaluated, including a PEGylated fibrin gel. PEGylation with a difunctional PEG derivative retarded fibrinogen migration during through sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) as a result of crosslinking, similarly, degradation was slowed in the PEGylated gel. ES cell proliferation was higher in both the fibrin and PEGylated fibrin gels versus 2D and methylcellulose controls. FACS analysis and real-time-PCR revealed differences in patterns of differentiation for the various culture systems. Culture in PEGylated fibrin or methylcellulose culture demonstrated features characteristic of less extensive differentiation relative to fibrin and 2D culture as evidenced by the transcription factor Oct-4. Fibrin gels showed gene and protein expression similar to that in 2D culture. Both fibrin and 2D cultures demonstrated statistically greater cell numbers positive for the vascular mesoderm marker, VE-cadherin.
Mesh Headings (Keywords): Animals, Biocompatible Materials, Cell Culture Techniques, Cell Differentiation, Cell Line, Cell Proliferation, Cell Survival, Cells, Cultured, Fibrin, Mice, Stem Cells, Tissue Engineering
Check for Full Text / PubMed Unique Identifier (PMID): 16860386
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