Monoclonal mice generated by nuclear transfer from mature B and T donor cells

Abstract

Cloning from somatic cells is inefficient, with most clones dying during gestation1,2. Cloning from embryonic stem (ES) cells is much more effective, suggesting that the nucleus of an embryonic cell is easier to reprogram3,4,5,6,7. It is thus possible that most surviving clones are, in fact, derived from the nuclei of rare somatic stem cells present in adult tissues, rather than from the nuclei of differentiated cells, as has been assumed1,8,9. Here we report the generation of monoclonal mice by nuclear transfer from mature lymphocytes. In a modified two-step cloning procedure, we established ES cells from cloned blastocysts and injected them into tetraploid blastocysts to generate mice. In this approach, the embryo is derived from the ES cells and the extra-embryonic tissues from the tetraploid host6. Animals cloned from a B-cell nucleus were viable and carried fully rearranged immunoglobulin alleles in all tissues. Similarly, a mouse cloned from a T-cell nucleus carried rearranged T-cell-receptor genes in all tissues. This is an unequivocal demonstration that a terminally differentiated cell can be reprogrammed to produce an adult cloned animal.

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Figure 1: Derivation of ES cells and monoclonal mice by nuclear transfer from mature B and T lymphocytes.
Figure 2: Immunological analyses of B- and T-cell-derived mice.

References

  1. 1

    Rideout, W. M. III, Eggan, K. & Jaenisch, R. Nuclear cloning and epigenetic reprogramming of the genome. Science 293, 1093–1098 (2001)

    CAS  Article  Google Scholar 

  2. 2

    Solter, D. Mammalian cloning: advances and limitations. Nature Rev. Genet. 1, 199–207 (2000)

    CAS  Article  Google Scholar 

  3. 3

    Wakayama, T., Rodriguez, I., Perry, A. C., Yanagimachi, R. & Mombaerts, P. Mice cloned from embryonic stem cells. Proc. Natl Acad. Sci. USA 96, 14984–14989 (1999)

    ADS  CAS  Article  Google Scholar 

  4. 4

    Rideout, W. M. et al. Generation of mice from wild-type and targeted ES cells by nuclear cloning. Nature Genet. 24, 109–110 (2000)

    CAS  Article  Google Scholar 

  5. 5

    Humpherys, D. et al. Epigenetic instability in ES cells and cloned mice. Science 293, 95–97 (2001)

    CAS  Article  Google Scholar 

  6. 6

    Eggan, K. et al. Hybrid vigor, fetal overgrowth, and viability of mice derived by nuclear cloning and tetraploid embryo complementation. Proc. Natl Acad. Sci. USA 98, 6209–6214 (2001)

    ADS  CAS  Article  Google Scholar 

  7. 7

    Wakayama, T. & Yanagimachi, R. Mouse cloning with nucleus donor cells of different age and type. Mol. Reprod. Dev. 58, 376–383 (2001)

    CAS  Article  Google Scholar 

  8. 8

    Weissman, I. L. Stem cells: units of development, units of regeneration, and units in evolution. Cell 100, 157–168 (2000)

    CAS  Article  Google Scholar 

  9. 9

    Liu, L. Cloning efficiency and differentiation. Nature Biotechnol. 19, 406 (2001)

    Article  Google Scholar 

  10. 10

    Gurdon, J., Laskey, R. & Reeves, O. The developmental capacity of nuclei transplanted from keratinized skin cells of adult frogs. J. Embryol. Exp. Morphol. 34, 93–112 (1975)

    CAS  PubMed  Google Scholar 

  11. 11

    DiBerardino, M. Genetic stability and modulation of metazoan nuclei transplanted into eggs and oocytes. Differentiation 17, 17–30 (1980)

    CAS  Article  Google Scholar 

  12. 12

    Briggs, R. & King, T. Changes in the nuclei of differentiating endoderm cells as revealed by nuclear transplantation. J. Morphol. 100, 269–311 (1957)

    Article  Google Scholar 

  13. 13

    McCreath, K. J. et al. Production of gene-targeted sheep by nuclear transfer from cultured somatic cells. Nature 405, 1066–1069 (2000)

    ADS  CAS  Article  Google Scholar 

  14. 14

    Wabl, M. R., Brun, R. B. & Du Pasquier, L. Lymphocytes of the toad Xenopus laevis have the gene set for promoting tadpole development. Science 190, 1310–1312 (1975)

    ADS  CAS  Article  Google Scholar 

  15. 15

    Wakayama, T., Perry, A. C., Zuccotti, M., Johnson, K. R. & Yanagimachi, R. Full-term development of mice from enucleated oocytes injected with cumulus cell nuclei. Nature 394, 369–374 (1998)

    ADS  CAS  Article  Google Scholar 

  16. 16

    Galli, C. Mammalian leukocytes contain all the genetic information necessary for the development of a new individual. Cloning 1, 161–170 (1999)

    CAS  Article  Google Scholar 

  17. 17

    Rajewsky, K. Clonal selection and learning in the antibody system. Nature 381, 751–758 (1996)

    ADS  CAS  Article  Google Scholar 

  18. 18

    Kisielow, P. & von Boehmer, H. Development and selection of T cells: facts and puzzles. Adv. Immunol. 58, 87–209 (1995)

    CAS  Article  Google Scholar 

  19. 19

    Nagy, A., Rossant, J., Nagy, R., Abramow-Newerly, W. & Roder, J. C. Derivation of completely cell culture-derived mice from early-passage embryonic stem cells. Proc. Natl Acad. Sci. USA 90, 8424–8428 (1993)

    ADS  CAS  Article  Google Scholar 

  20. 20

    Kawase, E., Yamazaki, Y., Yagi, T., Yanagimachi, R. & Pedersen, R. A. Mouse embryonic stem (ES) cell lines established from neuronal cell-derived cloned blastocysts. Genesis 28, 156–163 (2000)

    CAS  Article  Google Scholar 

  21. 21

    Munsie, M. J. et al. Isolation of pluripotent embryonic stem cells from reprogrammed adult mouse somatic cell nuclei. Curr. Biol. 10, 989–992 (2000)

    CAS  Article  Google Scholar 

  22. 22

    Wakayama, T. et al. Differentiation of embryonic stem cell lines generated from adult somatic cells by nuclear transfer. Science 292, 740–743 (2001)

    ADS  CAS  Article  Google Scholar 

  23. 23

    Kouskoff, V. & Nemazee, D. Role of receptor editing and revision in shaping the B and T lymphocyte repertoire. Life Sci. 69, 1105–1113 (2001)

    CAS  Article  Google Scholar 

  24. 24

    Chen, J., Lansford, R., Stewart, V., Young, F. & Alt, F. W. RAG-2-deficient blastocyst complementation: an assay of gene function in lymphocyte development. Proc. Natl Acad. Sci. USA 90, 4528–4532 (1993)

    ADS  CAS  Article  Google Scholar 

  25. 25

    Cascalho, M., Ma, A., Lee, S., Masat, L. & Wabl, M. A quasi-monoclonal mouse. Science 272, 1649–1652 (1996)

    ADS  CAS  Article  Google Scholar 

  26. 26

    Wells, D. N., Misica, P. M. & Tervit, H. R. Production of cloned calves following nuclear transfer with cultured adult mural granulosa cells. Biol. Reprod. 60, 996–1005 (1999)

    CAS  Article  Google Scholar 

  27. 27

    Chess, A., Simon, I., Cedar, H. & Axel, R. Allelic inactivation regulates olfactory receptor gene expression. Cell 78, 823–834 (1994)

    CAS  Article  Google Scholar 

  28. 28

    Livak, F., Petrie, H. T., Crispe, I. N. & Schatz, D. G. In-frame TCRδ gene rearrangements play a critical role in the αβ/γδ T cell lineage decision. Immunity 2, 617–627 (1995)

    CAS  Article  Google Scholar 

  29. 29

    Whitehurst, C. E., Chattopadhyay, S. & Chen, J. Control of V(D)J recombinational accessibility of the Dβ1 gene segment at the TCRβ locus by a germline promoter. Immunity 10, 313–322 (1999)

    CAS  Article  Google Scholar 

  30. 30

    Novobrantseva, T. I. et al. Rearrangement and expression of immunoglobulin light chain genes can precede heavy chain expression during normal B cell development in mice. J. Exp. Med. 189, 75–88 (1999)

    CAS  Article  Google Scholar 

Download references

Acknowledgements

We thank W. Rideout, K. Eggan, D. Humpherys and Z. Wang for discussions on the project; S. Nguyen, V. Haase, T. Novobrantseva and C. J. Ryu for advice with immunological analyses and for Southern blot probes; J. Dausman and R. Flannery for animal care; and A. Chess, C. Beard and J. Gribnau for critical reading of the manuscript. K.H. was supported by a PhD fellowship from the Boehringer Ingelheim Fonds. Support to R.J. was from the National Cancer Institute.

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Correspondence to Rudolf Jaenisch.

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Hochedlinger, K., Jaenisch, R. Monoclonal mice generated by nuclear transfer from mature B and T donor cells. Nature 415, 1035–1038 (2002). https://doi.org/10.1038/nature718

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