Generation of Novel Rat and Human Pluripotent Stem Cells by Reprogramming and Chemical Approaches

  • Wenlin Li
  • Sheng DingEmail author
Part of the Methods in Molecular Biology book series (MIMB, volume 636)


Although embryonic stem cells (ESCs) have been established from mice since 1981, attempts to derive its counterparts from various other mammals, including rats, have not succeeded. Recently, induced pluripotent stem cells (iPSCs) have been generated from both mouse and human somatic cells by genetic transduction. We had successfully established novel rat iPSCs (riPSCs), which can be homogenously maintained by LIF and a cocktail of ALK5 inhibitor, GSK3 inhibitor and MEK inhibitor. riPSCs share conventional mouse ESC characteristics and most importantly can contribute extensively to chimeras. We also generated novel human iPSCs (hiPSCs) with “mouse ESC-like” characteristics, which can be surprisingly maintained in culture in the presence of MEK inhibitor and ALK5 inhibitor.

Key words

Embryonic stem cells Pluripotent stem cells iPS cells Inhibitors 


  1. 1.
    Tesar, P. J., Chenoweth, J. G., Brook, F. A., Davies, T. J., Evans, E. P., Mack, D. L., et al. (2007) New cell lines from mouse epiblast share defining features with human embryonic stem cells. Nature. 448, 196–199.CrossRefPubMedGoogle Scholar
  2. 2.
    Brons, I. G., Smithers, L. E., Trotter, M. W., Rugg-Gunn, P., Sun, B., Chuva de Sousa Lopes, S. M., et al. (2007) Derivation of pluripotent epiblast stem cells from mammalian embryos. Nature. 448, 191–195.CrossRefPubMedGoogle Scholar
  3. 3.
    Niwa, H., Burdon, T., Chambers, I., and Smith, A. (1998) Self-renewal of pluripotent embryonic stem cells is mediated via activation of STAT3. Genes Dev. 12, 2048–2060.CrossRefPubMedGoogle Scholar
  4. 4.
    Ying, Q. L., Nichols, J., Chambers, I., and Smith, A. (2003) BMP induction of Id proteins suppresses differentiation and sustains embryonic stem cell self-renewal in collaboration with STAT3. Cell. 115, 281–292.CrossRefPubMedGoogle Scholar
  5. 5.
    Qi, X., Li, T. G., Hao, J., Hu, J., Wang, J., Simmons, H., et al. (2004) BMP4 supports self-renewal of embryonic stem cells by inhibiting mitogen-activated protein kinase pathways. Proc Natl Acad Sci USA. 101, 6027–6032.CrossRefPubMedGoogle Scholar
  6. 6.
    Dvorak, P., Dvorakova, D., Koskova, S., Vodinska, M., Najvirtova, M., Krekac, D., et al. (2005) Expression and potential role of fibroblast growth factor 2 and its receptors in human embryonic stem cells. Stem Cells. 23, 1200–1211.CrossRefPubMedGoogle Scholar
  7. 7.
    Xu, R. H., Chen, X., Li, D. S., Li, R., Addicks, G. C., Glennon, C., et al. (2002) BMP4 initiates human embryonic stem cell differentiation to trophoblast. Nat Biotechnol. 20, 1261–1264.CrossRefPubMedGoogle Scholar
  8. 8.
    Sato, N., Sanjuan, I. M., Heke, M., Uchida, M., Naef, F., and Brivanlou, A. H. (2003) Molecular signature of human embryonic stem cells and its comparison with the mouse. Dev Biol. 260, 404–413.CrossRefPubMedGoogle Scholar
  9. 9.
    Martin, G. R. (1981) Isolation of a pluripotent cell line from early mouse embryos cultured in medium conditioned by teratocarcinoma stem cells. Proc Natl Acad Sci USA. 78, 7634–7638.CrossRefPubMedGoogle Scholar
  10. 10.
    Ueda, S., Kawamata, M., Teratani, T., Shimizu, T., Tamai, Y., Ogawa, H., et al. (2008) Establishment of rat embryonic stem cells and making of chimera rats. PLoS ONE. 3, e2800.CrossRefPubMedGoogle Scholar
  11. 11.
    Demers, S. P., Yoo, J. G., Lian, L., Therrien, J., and Smith, L. C. (2007) Rat embryonic stem-like (ES-like) cells can contribute to extraembryonic tissues in vivo. Cloning Stem Cells. 9, 512–522.CrossRefPubMedGoogle Scholar
  12. 12.
    Schulze, M., Ungefroren, H., Bader, M., and Fandrich, F. (2006) Derivation, maintenance, and characterization of rat embryonic stem cells in vitro. Methods Mol Biol. 329, 45–58.PubMedGoogle Scholar
  13. 13.
    Ruhnke, M., Ungefroren, H., Zehle, G., Bader, M., Kremer, B., and Fandrich, F. (2003) Long-term culture and differentiation of rat embryonic stem cell-like cells into neuronal, glial, endothelial, and hepatic lineages. Stem Cells. 21, 428–436.CrossRefPubMedGoogle Scholar
  14. 14.
    Takahashi, K., Tanabe, K., Ohnuki, M., Narita, M., Ichisaka, T., Tomoda, K., et al. (2007) Induction of pluripotent stem cells from adult human fibroblasts by defined factors. Cell. 131, 861–872.CrossRefPubMedGoogle Scholar
  15. 15.
    Yu, J., Vodyanik, M. A., Smuga-Otto, K., Antosiewicz-Bourget, J., Frane, J. L., Tian, S., et al. (2007) Induced pluripotent stem cell lines derived from human somatic cells. Science. 318, 1917–1920.CrossRefPubMedGoogle Scholar
  16. 16.
    Takahashi, K., and Yamanaka, S. (2006) Induction of pluripotent stem cells from mouse embryonic and adult fibroblast cultures by defined factors. Cell. 126, 663–676.CrossRefPubMedGoogle Scholar
  17. 17.
    Dimos, J. T., Rodolfa, K. T., Niakan, K. K., Weisenthal, L. M., Mitsumoto, H., Chung, W., et al. (2008) Induced pluripotent stem cells generated from patients with ALS can be differentiated into motor neurons. Science. 321, 1218–1221.CrossRefPubMedGoogle Scholar
  18. 18.
    Han, J., and Sidhu, K. S. (2008) Current concepts in reprogramming somatic cells to pluripotent state. Curr Stem Cell Res Ther. 3, 66–74.CrossRefPubMedGoogle Scholar
  19. 19.
    Park, I. H., Zhao, R., West, J. A., Yabuuchi, A., Huo, H., Ince, T. A., et al. (2008) Reprogramming of human somatic cells to pluripotency with defined factors. Nature. 451, 141–146.CrossRefPubMedGoogle Scholar

Copyright information

© Springer Science+Business Media, LLC 2010

Authors and Affiliations

  1. 1.Department of ChemistryThe Scripps Research InstituteLa JollaUSA

Personalised recommendations