The rat represents an animal model highly attractive for studying pharmacology, physiology, aging, cardiovascular diseases, etc., that in many aspects is more adequate than the mouse model. Derivation of induced pluripotent stem cells from rats (riPS) opens the opportunity for gene targeting in specific rat strains, as well as for the development of new protocols for the treatment of different degenerative diseases. Here we report an improved protocol for riPS cell generation, which is based on lentivirus delivery of reprogramming factors with their subsequent excision from the genome, application of serum-free media and chemical inhibitors MEK and GSK. We compared various conditions for riPS cell derivation, analyzed the cell karyotype, and assessed the pluripotency of the established cells. These data may prompt further iPS cell-based gene targeting in rat, as well as the development of iPS-based cell therapy, using this animal model.
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- iPS cells:
induced pluripotent stem cells
mouse embryonic fibroblasts
- ES cells:
embryonic stem cells
enhanced green fluorescent protein
glycogen synthase kinase
leukemia inhibitory factor
phosphate buffer saline
Aoi, T., Yae, K., Nakagawa, M., Ichisaka, T., Okita, K., Takahashi, K., Chiba, T., and Yamanaka, S., Generation of Pluripotent Stem Cells from Adult Mouse Liver and Stomach Cells, Science, 2008, vol. 321, pp. 699–702.
Brambrink, T., Foreman, R., Welstead, G.G., Lengner, C.J., Wernig, M., Suh, H., and Jaenisch, R., Sequential Expression of Pluripotency Markers during Direct Reprogramming of Mouse Somatic Cells, Cell Stem Cell, 2008, vol. 2, pp. 151–159.
Buehr, M., Meek, S., Blair, K., Yang, J., Ure, J., Silva, J., McLay, R., Hall, J., Ying, Q.L., and Smith, A., Capture of Authentic Embryonic Stem Cells from Rat Blastocysts, Cell, 2008, vol. 135, pp. 1287–1298.
Evans, M.J. and Kaufman, M.H., Establishment in Culture of Pluripotential Cells from Mouse Embryos, Nature, 1981, vol. 292, pp. 154–156.
Hanna, J., Markoulaki, S., Schorderet, P., Carey, B.W., Beard, C., et al., Direct Reprogramming of Terminally Differentiated Mature B Lymphocytes to Pluripotency, Cell, 2008, vol. 133, pp. 250–264.
Holubcová, Z., Matula, P., Sedláčková, M., Vinarský, V., Doležalová, D., Bárta, T., Dvořák, P., and Hampl, A., Human Embryonic Stem Cells Suffer from Centrosomal Amplification, Stem Cells, 2010, vol. 29, pp. 46–56.
Li, W., Wei, W., Zhu, S., Zhu, J., Shi, Y., Lin, T., Hao, E., Hayek, A., Deng, H., and Ding, S., Generation of Rat and Human Induced Pluripotent Stem Cells by Combining Genetic Reprogramming and Chemical Inhibitors, Cell Stem Cell, 2009, vol. 4, pp. 16–19.
Liao, J., Cui, C., Chen, S., Ren, J., Chen, J., Gao, Y., Li, H., Jia, N., Cheng, L., Xiao, H., and Xiao, L., Generation of Induced Pluripotent Stem Cell Lines from Adult Rat Cells, Cell. Stem Cell, 2009, vol. 4, pp. 11–5.
Minina, Iu.M., Zhdanova, N.S., Shilov, A.G., Tolkunova, E.N., Liskovykh, M.A., and Tomilin, A.N., Chromosomal Instability of in vitro Cultured Mouse Embryonic Stem Cells and Induced Pluripotent Stem Cells, Tsitologiya, 2010, vol. 52, pp. 420–425.
O’Brien, S.J., Menninger, J.C., and Nash, W.G., Atlas of Mammalian Chromosomes, Hoboken, NJ: Wiley-Liss. 2006, vol. XLII.
Okita, K., Ichisaka, T., and Yamanaka, S., Generation of Germline-Competent Induced Pluripotent Stem Cells, Nature, 2007, vol. 448, pp. 313–317.
Silva, J., Barrandon, O., Nichols, J., Kawaguchi, J., Theunissen, T.W., and Smith, A., Promotion of Reprogramming to Ground State Pluripotency by Signal Inhibition, PLoS Biol., 2008, vol. 6, p. e253.
Stadtfeld, M., Brennand, K., and Hochedlinger, K., Reprogramming of Pancreatic β Cells into Induced Pluripotent Stem Cells, Curr. Biol., 2008, vol. 18, pp. 890–894.
Sugawara, A., Goto, K., Sotomaru, Y., Sofuni, T., and Ito, T., Current Status of Chromosomal Abnormalities in Mouse Embryonic Stem Cell Lines Used in Japan, Comp. Med., 2006, vol. 56, pp. 31–34.
Takahashi, K. and Yamanaka, S., Induction of Pluripotent Stem Cells from Mouse Embryonic and Adult Fibroblast Cultures by Defined Factors, Cell, 2006, vol. 126, pp. 663–676.
Wiznerowicz, M. and Trono, D., Conditional Suppression of Cellular Genes: Lentivirus Vector-Mediated Drug-Inducible RNA Interference, J. Virol., 2003, vol. 77, pp. 8957–8961.
Ying, Q.L., Wray, J., Nichols, J., Batlle-Morera, L., Doble, B., Woodgett, J., Cohen, P., and Smith, A., The Ground State of Embryonic Stem Cell Self-Renewal, Nature, 2008, vol. 453, pp. 519–523.
Yu, J., Vodyanik, M.A., Smuga-Otto, K., Antosiewicz-Bourget, J., Frane, J.L., Tian, S., Nie, J., Jonsdottir, G.A., Ruotti, V., Stewart, R., Slukvin, I.I., and Thomson, J.A., Induced Pluripotent Stem Cell Lines Derived from Human Somatic Cells, Science, 2007, vol. 318, pp. 1917–1920.
Original Russian Text © M.A. Liskovykh, I.A. Chuykin, A. Ranjan, D.A. Safina, E.N. Tolkunova, Yu.M. Minina, N.S. Zhdanova, P.A. Dyban, John J. Mullins, E.I. Kostyleva, E.V. Chikhirzhina, M. Bader, N. Alenina, A.N. Tomilin, 2011, published in Tsitologiya, 2011, Vol. 53, No. 12, pp. 939–945.
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Liskovykh, M.A., Chuykin, I.A., Ranjan, A. et al. Generation of rat-induced pluripotent stem cells: Reprogramming and culture medium. Cell Tiss. Biol. 6, 115–121 (2012). https://doi.org/10.1134/S1990519X12020095
- induced pluripotent stem cells