Developing immortal cell lines from Xenopus embryos, four novel cell lines derived from Xenopus tropicalis

© The Author(s), 2022. This article is distributed under the terms of the Creative Commons Attribution License. The definitive version was published in Gorbsky, G. J., Daum, J. R., Sapkota, H., Summala, K., Yoshida, H., Georgescu, C., Wren, J. D., Peshkin, L., & Horb, M. E. Developing immortal cell lines from Xenopus embryos, four novel cell lines derived from Xenopus tropicalis. Open Biology, 12(7), (2022): 220089, https://doi.org/10.1098/rsob.220089.
The diploid anuran Xenopus tropicalis has emerged as a key research model in cell and developmental biology. To enhance the usefulness of this species, we developed methods for generating immortal cell lines from Nigerian strain (NXR_1018, RRID:SCR_013731) X. tropicalis embryos. We generated 14 cell lines that were propagated for several months. We selected four morphologically distinct lines, XTN-6, XTN-8, XTN-10 and XTN-12 for further characterization. Karyotype analysis revealed that three of the lines, XTN-8, XTN-10 and XTN-12 were primarily diploid. XTN-6 cultures showed a consistent mixed population of diploid cells, cells with chromosome 8 trisomy, and cells containing a tetraploid content of chromosomes. The lines were propagated using conventional culture methods as adherent cultures at 30°C in a simple, diluted L-15 medium containing fetal bovine serum without use of a high CO2 incubator. Transcriptome analysis indicated that the four lines were distinct lineages. These methods will be useful in the generation of cell lines from normal and mutant strains of X. tropicalis as well as other species of Xenopus.
This work was supported by Whitman fellowships to G.J.G. from the Marine Biological Laboratory, by grant no. 1645105 to G.J.G. and MEH from the National Science Foundation and by grant no. P40OD010997 from the Office of the Director, National Institutes of Health. L.P. has been supported by grant no. R01HD073104 from the National Institute of Child Health and Development.