The undersigned hereby assign all rights, included but not limited to copyright, for this manuscript to CMB Association upon its submission for consideration to publication on Cellular and Molecular Biology. The rights assigned include, but are not limited to, the sole and exclusive rights to license, sell, subsequently assign, derive, distribute, display and reproduce this manuscript, in whole or in part, in any format, electronic or otherwise, including those in existence at the time this agreement was signed. The authors hereby warrant that they have not granted or assigned, and shall not grant or assign, the aforementioned rights to any other person, firm, organization, or other entity. All rights are automatically restored to authors if this manuscript is not accepted for publication.
Coculturing embryonic stem cells with damaged hepatocytes leads to restoration of damage and high frequency of fusion
Corresponding Author(s) : D Xu
Cellular and Molecular Biology,
Vol. 55 No. 4: General Papers
Abstract
Controversy surrounds issue of cell fusion as a repair mechanism whereby stem cells regenerate. To identify the ratio of fusion happens between stem cells and damaged cells, hepatic cells were damaged with 200μM H2O2 for 2 hr. Then, mouse ESCs were cocultured with damaged human hepatocytes. Fusion was detected directly by karyotyping after 48hr coculture as well as by Oct4 promoter drove GFP signal. Results showed that average ratio of fusion in undamaged control group was 0.031 while ratio of fusion in damaged group was 0.357, which was 10 times higher than fusion happened in the control group. Meanwhile, GFP signal indicated that fusion induced hepatic cells' Oct-4 reactivation. Fusion derived hybrid cells contained chromosomes from both parental cells. Most of the chromosomes were from damaged human hepatic cells. Activity of damage-related enzymes LDH, SGOT and SGPT were significantly lower at 48hr coculture than at 12hr coculture. Expression of albumin in co-culture system was up-regulated after coculture, which indicated the reparation of damage after coculturing. Also, by applying RT-PCR and immunocytochemistry differentiation status of ES cells were evaluated. It was shown that ES cells differentiated to hepatic lineage cells and expressed hepatic genes and proteins.
Keywords
Hybrid cells
coculture
karyotype
differentiation.
Xu, D., Wang, F., Pan, Z., & Guo, Q. (2009). Coculturing embryonic stem cells with damaged hepatocytes leads to restoration of damage and high frequency of fusion. Cellular and Molecular Biology, 55(4), 1186–99. Retrieved from https://cellmolbiol.org/index.php/CMB/article/view/1061
Download Citation
Endnote/Zotero/Mendeley (RIS)BibTeX