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Editors Selection IGR 14-4
Animal Models: Stem cells
Comment by Yvonne Ou on:
51896 Generating embryonic stem cells from the inbred mouse strain DBA/2J, a model of glaucoma and other complex diseases, Reinholdt LG; Howell GR; Czechanski AM et al., PLoS ONE, 2012; 7: e50081
Over the past 20 years, the application of gene targeting technology in embryonic stem (ES) cells has led to the elucidation of the function of more than 5,000 mammalian genes and the creation of more than 500 models of human diseases. Furthermore, the function of the specific gene of interest can be analyzed in a temporal and spatial manner. Currently, the vast majority of the resources needed to generate these mouse lines have been developed in a limited number of strains, including the C57BL/6J and 129Sv/ImJ. These resources, however, are lacking for inbred mouse strains such as the DBA/2J. The improved ability to genetically manipulate genes of interest on a DBA/2J background will greatly accelerate our understanding of important genes and mechanisms in glaucoma pathogenesis. The paper by Reinholdt et al. reports the first use of DBA/2J ES cells to generate a genetically engineered mouse strain. Previous attempts to derive DBA/2J ES cells have failed, but the authors surmised that several inhibitors of signaling pathways that promote early differentiation may be useful in improving ES cell generation from the DBA/2J strain, as has been shown in other strains recalcitrant to ES cell derivation. The authors found that serum-free media containing inhibitors of early differentiation (FGF receptor tyrosine kinase inhibitor, MEK kinase inhibitor, glycogen synthase kinase-3 inhibitor) and LIF promoted germ line competency and pluripotency better than other derivation protocols tested. Lastly, the authors used the DBA/2J ES cells to create a conditional knockout allele for endothelin- 2 on the DBA/2J background. This study was rigorous in its techniques, utilizing not only immunolabeling for pluripotency markers and quantification by flow cytometry, but also the gold standard test of pluripotency for mouse ES cells (contribution to the germ line of a chimeric animal). What will be even more interesting is utilizing this technology to understand the roles of specific genes hypothesized to play a role in glaucomatous neurodegeneration, since now we will be able to generate conditional alleles of specific genes of interest on the DBA/2J background.
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