Developmental, Behavioral, and Physiological Phenotype of Cloned Mice

  • Kellie L. K. Tamashiro
  • Randall R. Sakai
  • Yukiko Yamazaki
  • Teruhiko Wakayama
  • Ryuzo Yanagimachi
Part of the Advances in Experimental Medicine and Biology book series (volume 591)


Cloning from adult somatic cells has been successful in at least ten species. Although generating viable cloned mammals from adult cells is technically feasible, prenatal and perinatal mortality is high and live cloned offspring have had health problems. This chapter summarizes the health consequences of cloning in mice and discusses possible mechanisms through which these conditions may arise. These studies have further significance as other assisted reproductive techniques (ART) also involve some of the same procedures used in cloning, and there are some reports that offspring generated by ART display aberrant phenotypes as well. At the moment, the long-term consequences of mammalian cloning remain poorly characterized. Data available thus far suggest that we should use this technology with great caution until numerous questions are addressed and answered.


Telomere Length Imprint Gene Cumulus Cell Nuclear Transfer Somatic Cell Nuclear Transfer 
These keywords were added by machine and not by the authors. This process is experimental and the keywords may be updated as the learning algorithm improves.


Unable to display preview. Download preview PDF.

Unable to display preview. Download preview PDF.


  1. 1.
    McGrath J, Solter D. Inability of mouse blastomere nuclei transferred to enucleated zygotes to support development in vitro. Science 1984; 226(4680):1317–9.PubMedCrossRefGoogle Scholar
  2. 2.
    Wilmut I, Schnieke AE, McWhir J et al. Viable offspring derived from fetal and adult mammalian cells. Nature 1997; 385(6619):810–3.PubMedCrossRefGoogle Scholar
  3. 3.
    Kato Y, Tani T, Sotomaru Y et al. Eight calves cloned from somatic cells of a single adult. Science 1998; 282(5396):2095–8.PubMedCrossRefGoogle Scholar
  4. 4.
    Wakayama T, Perry AC, Zuccotti M et al. Full-term development of mice from enucleated oocytes injected with cumulus cell nuclei. Nature 1998; 394(6691):369–74.PubMedCrossRefGoogle Scholar
  5. 5.
    Polejaeva IA, Chen SH, Vaught TD et al. Cloned pigs produced by nuclear transfer from adult somatic cells. Nature 2000; 407(6800):86–90.PubMedCrossRefGoogle Scholar
  6. 6.
    Keefer CL, Keyston R, Lazaris A et al. Production of cloned goats after nuclear transfer using adult somatic cells. Biol Reprod 2002; 66(1):199–203.PubMedCrossRefGoogle Scholar
  7. 7.
    Shin T, Kraemer D, Pryor J et al. A cat cloned by nuclear transplantation. Nature 2002; 415(6874):859.PubMedCrossRefGoogle Scholar
  8. 8.
    Chesne P, Adenot PG, Viglietta C et al. Cloned rabbits produced by nuclear transfer from adult somatic cells. Nat Biotechnol 2002; 20(4):366–9.PubMedCrossRefGoogle Scholar
  9. 9.
    Galli C, Lagutina I, Crotti G et al. Pregnancy: A cloned horse born to its dam twin. Nature 2003; 424(6949):635.PubMedCrossRefGoogle Scholar
  10. 10.
    Zhou Q, Renard JP, Le Friec G et al. Generation of fertile cloned rats by regulating oocyte activation. Science 2003; 1088313.Google Scholar
  11. 11.
    Lee BC, Kim MK, Jang G et al. Dogs cloned from adult somatic cells. Nature 2005; 436(7051):641.PubMedCrossRefGoogle Scholar
  12. 12.
    McCreath KJ, Howcroft J, Campbell KH et al. Production of gene-targeted sheep by nuclear transfer from cultured somatic cells. Nature 2000; 405(6790):1066–9.PubMedCrossRefGoogle Scholar
  13. 13.
    Schnieke AE, Kind AJ, Ritchie WA et al. Human factor IX transgenic sheep produced by transfer of nuclei from transfected fetal fibroblasts. Science 1997; 278(5346):2130–3.PubMedCrossRefGoogle Scholar
  14. 14.
    Lai L, Kolber-Simonds D, Park KW et al. Production of alpha-1,3-galactosyltransferase knockout pigs by nuclear transfer cloning. Science 2002; 295(5557):1089–92.PubMedCrossRefGoogle Scholar
  15. 15.
    Wilmut I, Paterson L. Somatic cell nuclear transfer. Nature 2003; 13(6–10):303–7.Google Scholar
  16. 16.
    Holt WV, Pickard AR, Prather RS. Wildlife conservation and reproductive cloning. Reproduction 2004; 127(3):317–24.PubMedCrossRefGoogle Scholar
  17. 17.
    Westhusin ME, Burghardt RC, Ruglia JN et al. Potential for cloning dogs. J Reprod Fertil Suppl 2001; 57:287–93.PubMedGoogle Scholar
  18. 18.
    Westhusin M, Hinrichs K, Choi YH et al. Cloning companion animals (horses, cats, and dogs). Cloning Stem Cells 2003; 5(4):301–17.PubMedCrossRefGoogle Scholar
  19. 19.
    Vogel G. Reproductive biology Cloning: Could humans be next? Science 2001; 291(5505):808b–809.CrossRefGoogle Scholar
  20. 20.
    Schatten G, Prather R, Wilmut I. Cloning claim is science fiction, not science. Science 2003; 299(5605):344.PubMedCrossRefGoogle Scholar
  21. 21.
    Wilmut I, Beaujean N, de Sousa PA et al. Somatic cell nuclear transfer. Nature 2002; 419(6907):583–6.PubMedCrossRefGoogle Scholar
  22. 22.
    Young LE, Fernandes K, McEvoy TG et al. Epigenetic change in IGF2R is associated with fetal overgrowth after sheep embryo culture. Nat Genet 2001; 27(2):153–4.PubMedCrossRefGoogle Scholar
  23. 23.
    Boiani M, Eckardt S, Scholer HR et al. Oct4 distribution and level in mouse clones: Consequences for pluripotency. Genes Dev 2002; 16(10):1209–19.PubMedCrossRefGoogle Scholar
  24. 24.
    Tamashiro KL, Wakayama T, Akutsu H et al. Cloned mice have an obese phenotype not transmitted to their offspring. Nat Med 2002; 8(3):262–7.PubMedCrossRefGoogle Scholar
  25. 25.
    Ecker DJ, Stein P, Xu Z et al. Long-term effects of culture of preimplantation mouse embryos on behavior. Proc Natl Acad Sci USA 2004; 101(6):1595–600.PubMedCrossRefGoogle Scholar
  26. 26.
    Fernandez-Gonzalez R, Moreira P, Bilbao A et al. Long-term effect of in vitro culture of mouse embryos with serum on mRNA expression of imprinting genes, development, and behavior. Proc Natl Acad Sci USA 2004; 101(16):5880–5.PubMedCrossRefGoogle Scholar
  27. 27.
    Solter D. Mammalian cloning: Advances and limitations. Nat Rev Genet 2000; 1(3):199–207.PubMedCrossRefGoogle Scholar
  28. 28.
    Perry AC, Wakayama T. Untimely ends and new beginnings in mouse cloning. Nat Genet 2002; 30(3):243–4.PubMedCrossRefGoogle Scholar
  29. 29.
    Young LE, Fairburn HR. Improving the safety of embryo technologies: Possible role of genomic imprinting. Theriogenology 2000; 53(2):627–48.PubMedCrossRefGoogle Scholar
  30. 30.
    Wakayama T, Rodriguez I, Perry AC et al. Mice cloned from embryonic stem cells. Proc Natl Acad Sci USA 1999; 96(26):14984–9.PubMedCrossRefGoogle Scholar
  31. 31.
    Humpherys D, Eggan K, Akutsu H et al. Epigenetic instability in ES cells and cloned mice. Science 2001; 293(5527):95–7.PubMedCrossRefGoogle Scholar
  32. 32.
    Humpherys D, Eggan K, Akutsu H et al. Abnormal gene expression in cloned mice derived from embryonic stem cell and cumulus cell nuclei. Proc Natl Acad Sci USA 2002; 99(20):12889–94.PubMedCrossRefGoogle Scholar
  33. 33.
    Gao S, McGarry M, Ferrier T et al. Effect of cell confluence on production of cloned mice using an inbred embryonic stem cell line. Biol Reprod 2003; 68(2):595–603.PubMedCrossRefGoogle Scholar
  34. 34.
    Yamazaki Y, Makino H, Hamaguchi-Hamada K et al. Assessment of the developmental totipotency of neural cells in the cerebral cortex of mouse embryo by nuclear transfer. Proc Natl Acad Sci USA 2001; 98(24):14022–6.PubMedCrossRefGoogle Scholar
  35. 35.
    Wakayama T, Yanagimachi R. Cloning of male mice from adult tail-tip cells. Nat Genet 1999; 22(2):127–8.PubMedCrossRefGoogle Scholar
  36. 36.
    Wakayama T, Yanagimachi R. Cloning the laboratory mouse. Semin Cell Dev Biol 1999; 10(3):253–8.PubMedCrossRefGoogle Scholar
  37. 37.
    Ogura A, Inoue K, Ogonuki N et al. Production of male cloned mice from fresh, cultured, and cryopreserved immature Sertoli cells. Biol Reprod 2000; 62(6):1579–84.PubMedCrossRefGoogle Scholar
  38. 38.
    Ogura A, Inoue K, Takano K et al. Birth of mice after nuclear transfer by electrofusion using tail tip cells. Mol Reprod Dev 2000; 57(1):55–9.PubMedCrossRefGoogle Scholar
  39. 39.
    Kishikawa H, Wakayama T, Yanagimachi R. Comparison of oocyte-activating agents for mouse cloning. Cloning Stem Cells 1999; 1(3):153–159.Google Scholar
  40. 40.
    Wakayama T, Shinkai Y, Tamashiro KL et al. Cloning of mice to six generations. Nature 2000; 407(6802):318–9.PubMedCrossRefGoogle Scholar
  41. 41.
    Tanaka S, Oda M, Toyoshima Y et al. Placentomegaly in cloned mouse concepti caused by expansion of the spongiotrophoblast layer. Biol Reprod 2001; 65(6):1813–21.PubMedCrossRefGoogle Scholar
  42. 42.
    Ogura A, Inoue K, Ogonuki N et al. Phenotypic effects of somatic cell cloning in the mouse. Cloning Stem Cells 2002; 4(4):397–405.PubMedCrossRefGoogle Scholar
  43. 43.
    Singh U, Fohn LE, Wakayama T et al. Different molecular mechanisms underlie placental overgrowth phenotypes caused by interspecies hybridization, cloning, and Esx1 mutation. Dev Dyn 2004; 230(1):149–64.PubMedCrossRefGoogle Scholar
  44. 44.
    Cibelli JB, Stice SL, Golueke PJ et al. Cloned transgenic calves produced from nonquiescent fetal fibroblasts. Science 1998; 280(5367):1256–8.PubMedCrossRefGoogle Scholar
  45. 45.
    Ohgane J, Wakayama T, Senda S et al. The Sall3 locus is an epigenetic hotspot of aberrant DNA methylation associated with placentomegaly of cloned mice. Genes Cells 2004; 9(3):253–60.PubMedCrossRefGoogle Scholar
  46. 46.
    Inoue K, Kohda T, Lee J et al. Faithful expression of imprinted genes in cloned mice. Science 2002; 295(5553):297.PubMedCrossRefGoogle Scholar
  47. 47.
    Ohgane J, Wakayama T, Kogo Y et al. DNA methylation variation in cloned mice. Genesis 2001; 30(2):45–50.PubMedCrossRefGoogle Scholar
  48. 48.
    Mann MR, Chung YG, Nolen LD et al. Disruption of imprinted gene methylation and expression in cloned preimplantation stage mouse embryos. Biol Reprod 2003; 69(3):902–14.PubMedCrossRefGoogle Scholar
  49. 49.
    Eggan K, Akutsu H, Loring J et al. Hybrid vigor, fetal overgrowth, and viability of mice derived by nuclear cloning and tetraploid embryo complementation. Proc Natl Acad Sci USA 2001; 98(11):62O9–14.CrossRefGoogle Scholar
  50. 50.
    Shimozawa N, Ono Y, Kimoto S et al. Abnormalities in cloned mice are not transmitted to the progeny. Genesis 2002; 34(3):203–7.PubMedCrossRefGoogle Scholar
  51. 51.
    Tamashiro KL, Sakai RR, Yamazaki Y et al. Health consequences of cloning mice. Health Consequences of Cloning: A Inui, Taylor and Francis Books. 2005:1–16.Google Scholar
  52. 52.
    Young LE, Sinclair KD, Wilmut I. Large offspring syndrome in cattle and sheep. Rev Reprod 1998; 3(3):155–63.PubMedCrossRefGoogle Scholar
  53. 53.
    Fox WM. Reflex-ontogeny and behavioural development of the mouse. Anim Behav 1965; 13(2):234–41.PubMedCrossRefGoogle Scholar
  54. 54.
    Tamashiro KL, Wakayama T, Blanchard RJ et al. Postnatal growth and behavioral development of mice cloned from adult cumulus cells. Biol Reprod 2000; 63(1):328–34.PubMedCrossRefGoogle Scholar
  55. 55.
    Morris RGM. Spatial localization does not require the presence of local cues. Learn Motiv 1981; 12:239–260.CrossRefGoogle Scholar
  56. 56.
    Shiels PG, Kind AJ, Campbell KH et al. Analysis of telomere lengths in cloned sheep. Nature 1999; 399(6734):316–7.PubMedCrossRefGoogle Scholar
  57. 57.
    Kuhholzer-Cabot B, Brem G. Aging of animals produced by somatic cell nuclear transfer. Exp Gerontol 2002; 37(12):1317–23.PubMedCrossRefGoogle Scholar
  58. 58.
    Ogonuki N, Inoue K, Yamamoto Y et al. Early death of mice cloned from somatic cells. Nat Genet 2002; 30(3):253–4.PubMedCrossRefGoogle Scholar
  59. 59.
    Shay JW, Wright WE. Hayflick, his limit, and cellular ageing. Nat Rev Mol Cell Biol 2000; 1(1):72–6.PubMedCrossRefGoogle Scholar
  60. 60.
    Tian XC, Xu J, Yang X. Normal telomere lengths found in cloned cattle. Nat Genet 2000; 26(3):272–3.PubMedCrossRefGoogle Scholar
  61. 61.
    Kubota C, Tian XC, Yang X. Serial bull cloning by somatic cell nuclear transfer. Nat Biotechnol 2004; 22:693–694.PubMedCrossRefGoogle Scholar
  62. 62.
    Lanza RP, Cibelli JB, Blackwell C et al. Extension of cell life-span and telomere length in animals cloned from senescent somatic cells. Science 2000; 288(5466):665–9.PubMedCrossRefGoogle Scholar
  63. 63.
    Jiang L, Carter B, Xu J et al. Telomere lengths in cloned transgenic pigs. Biol Reprod 2004.Google Scholar
  64. 64.
    Tamashiro KL, Wakayama T, Yamazaki Y et al. Phenotype of cloned mice: Development, behavior and physiology. Experimental Biology and Medicine 2003; 228(10):1193–1200.PubMedGoogle Scholar
  65. 65.
    Renard JP, Chastant S, Chesne P et al. Lymphoid hypoplasia and somatic cloning. Lancet 1999; 353(9163):1489–91.PubMedCrossRefGoogle Scholar
  66. 66.
    Keefer CL, Baldassarre H, Keyston R et al. Generation of dwarf goat (Capra hircus) clones following nuclear transfer with transfected and nontransfected fetal fibroblasts and in vitro-matured oocytes. Biol Reprod 2001; 64(3):849–56.PubMedCrossRefGoogle Scholar
  67. 67.
    Blasco MA. Immunosenescence phenotypes in the telomerase knockout mouse. Springer Semin Immunopathol 2002; 24(1):5–85.CrossRefGoogle Scholar
  68. 68.
    Betts D, Bordignon V, Hill J et al. Reprogramming of telomerase activity and rebuilding of telomere length in cloned cattle. Proc Natl Acad Sci USA 2001; 98(3):1077–82.PubMedCrossRefGoogle Scholar
  69. 69.
    Miyashita N, Shiga K, Yonai M et al. Remarkable differences in telomere lengths among cloned cattle derived from different cell types. Biol Reprod 2002; 66(6):1649–55.PubMedCrossRefGoogle Scholar
  70. 70.
    Aldhous P. The cloned mouse that roared is silenced. Nature 2000; 405:268.CrossRefGoogle Scholar

Copyright information

© Landes Bioscience and Springer Science+Business Media 2007

Authors and Affiliations

  • Kellie L. K. Tamashiro
    • 1
    • 2
  • Randall R. Sakai
    • 2
  • Yukiko Yamazaki
    • 3
  • Teruhiko Wakayama
    • 3
    • 4
  • Ryuzo Yanagimachi
    • 3
  1. 1.Department of Psychiatry and Behavioral SciencesJohns Hopkins University, School of MedicineBaltimoreUSA
  2. 2.Neuroscience Program, Department of PsychiatryUniversity of Cincinnati, College of MedicineCincinnatiUSA
  3. 3.Institute for Biogenesis ResearchUniversity of Hawaii School of MedicineHonoluluUSA
  4. 4.Laboratory for Genomic Reprogramming, Center for Developmental BiologyRIKEN KobeKobeJapan

Personalised recommendations