Transdifferentiation of Fibroblast into Chondrocytes
Date
2015-01-12
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Abstract
Great strides have been made in generating autologous induced pluripotent stem cells (iPSCs) via the process of cellular reprogramming. A key step in cellular reprogramming is the opening of chromatin. In newer transdifferentiation schemes, instead of reverting cells all the way to the embryonic state, cells can instead be redirected to specific cell types. One approach recently applied to the generation of cardiac and neural tissue involves partially reprogramming fibroblasts by blocking the JAK-STAT pathway followed by the introduction of specific differentiation factors. We have used a similar approach to transdifferentiate mouse embryo fibroblasts into chondrocytes. Following one day of exposure to the transcription factors Oct4, Sox2, Klf4 and c-Myc, (OSKM factors) and 10 days of exposure to a Jak inhibitor, resulting cells are dissociated and used to form micromass cultures for chondrogenic differentiation in the presence of TGF-b1 , and BMP-2. Cells formed aggregates, which differentiated into chondrocytes within 15 days, eventually formed hyaline and hypertrophic cartilage tissue based upon Alcian Blue, safranin-o and hematoxylin and eosin (H&E) staining. RT-PCR was used to establish a timeline of chondrocyte/cartilage gene expression. We also observed the expression of Col10 and Mmp13, which suggested the development of bone. Jak inhibitor made in all cases transdifferentiation into cartilage more efficient by avoiding bone formation.
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Education--Sciences, Biology--Cell, Biology--Molecular
Citation
Cota-Elizondo, P. A. (2015). Transdifferentiation of Fibroblast into Chondrocytes (Master's thesis, University of Calgary, Calgary, Canada). Retrieved from https://prism.ucalgary.ca. doi:10.11575/PRISM/28508