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Membrane Culture and Reduced Oxygen Tension Enhances Chondrogenesis of Equine Cord Blood Mesenchymal Stromal Cells in Vitro

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Title: Membrane Culture and Reduced Oxygen Tension Enhances Chondrogenesis of Equine Cord Blood Mesenchymal Stromal Cells in Vitro
Author: Co, Carmon
Department: Department of Biomedical Sciences
Program: Biomedical Sciences
Advisor: Koch, Thomas
Abstract: Variations of culture conditions impact the quality of neocartilage generated in vitro from mesenchymal stromal cells (MSC). The objective of this study was to evaluate and compare the effect of reduced oxygen and membrane culture during in vitro chondrogenesis of equine cord blood (CB) MSC. CB-MSC (n=5 horses) were expanded at 21% oxygen prior to 3 week differentiation in membrane or pellet culture at 5% and 21% oxygen. Assessment included histological examination (H&E, Toluidine Blue, Immunohistochemistry for type I and II collagen), protein quantification by Hydroxyproline assay and Dimethylmethylene assay, and mRNA analysis for collagen IA1, collagen IIA1, collagen XA1, HIF1α and Sox9. The combination of reduced oxygen and membrane culture produced neocartilage that most closely resembled hyaline cartilage. Membrane culture resulted in increased wet mass, homogenous matrix morphology and an increase in total collagen content, while 5% oxygen culture resulted in higher GAG and type II collagen content.
URI: http://hdl.handle.net/10214/7690
Date: 2013-12
Rights: Attribution-NonCommercial-NoDerivs 2.5 Canada


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Attribution-NonCommercial-NoDerivs 2.5 Canada Except where otherwise noted, this item's license is described as Attribution-NonCommercial-NoDerivs 2.5 Canada