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Science 22 May 1998:
Vol. 280. no. 5367, pp. 1256 - 1258
DOI: 10.1126/science.280.5367.1256
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Reports

Cloned Transgenic Calves Produced from Nonquiescent Fetal Fibroblasts

Jose B. Cibelli, Steve L. Stice, Paul J. Golueke, Jeff J. Kane, Joseph Jerry, Cathy Blackwell, F. Abel Ponce de León, James M. Robl *

An efficient system for genetic modification and large-scale cloning of cattle is of importance for agriculture, biotechnology, and human medicine. Here, actively dividing fetal fibroblasts were genetically modified with a marker gene, a clonal line was selected, and the cells were fused to enucleated mature oocytes. Out of 28 embryos transferred to 11 recipient cows, three healthy, identical, transgenic calves were generated. Furthermore, the life-span of near senescent fibroblasts could be extended by nuclear transfer, as indicated by population doublings in fibroblast lines derived from a 40-day-old fetal clone. With the ability to extend the life-span of these primary cultured cells, this system would be useful for inducing complex genetic modifications in cattle.

J. B. Cibelli, J. Jerry, J. M. Robl, Department of Veterinary and Animal Sciences, University of Massachusetts, Amherst, MA 01003, USA.
S. L. Stice, P. J. Golueke, J. J. Kane, C. Blackwell, Advanced Cell Technology, Incorporated, One Innovation Drive, Worcester, MA 01605, USA.
F. A. Ponce de León, Animal Sciences, College of Agricultural, Food and Environmental Sciences, 1404 Gortner Avenue, St Paul, MN 55108, USA.
*   To whom correspondence should be addressed. E-mail: robl{at}vasci.umass.edu

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Fate of Donor Mitochondrial DNA in Cloned Bovine Embryos Produced by Microinjection of Cumulus Cells.
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Dependence of DNA Synthesis and In Vitro Development of Bovine Nuclear Transfer Embryos on the Stage of the Cell Cycle of Donor Cells and Recipient Cytoplasts.
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Production of Cloned Cattle from In Vitro Systems.
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Clinical, Hormonal, and Hematologic Characteristics of Bovine Calves Derived from Nuclei from Somatic Cells.
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In Vitro Development of Horse Oocytes Reconstructed with the Nuclei of Fetal and Adult Cells.
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Effect of Fibroblast Donor Cell Age and Cell Cycle on Development of Bovine Nuclear Transfer Embryos In Vitro.
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PNAS
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Cloned Mice from Fetal Fibroblast Cells Arrested at Metaphase by a Serial Nuclear Transfer.
Y. Ono, N. Shimozawa, M. Ito, and T. Kono (2001)
Biol Reprod 64, 44-50
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Development of Cloned Embryos from Adult Rabbit Fibroblasts: Effect of Activation Treatment and Donor Cell Preparation.
A. Dinnyés, Y. Dai, M. Barber, L. Liu, J. Xu, P. Zhou, and X. Yang (2001)
Biol Reprod 64, 257-263
   Abstract »    Full Text »
Direct Exposure of Chromosomes to Nonactivated Ovum Cytoplasm Is Effective for Bovine Somatic Cell Nucleus Reprogramming.
T. Tani, Y. Kato, and Y. Tsunoda (2001)
Biol Reprod 64, 324-330
   Abstract »    Full Text »
The Ethical Validity of Using Nuclear Transfer in Human Transplantation.
R. P. Lanza, A. L. Caplan, L. M. Silver, J. B. Cibelli, M. D. West, and R. M. Green (2000)
JAMA 284, 3175-3179
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Evidence for Placental Abnormality as the Major Cause of Mortality in First-Trimester Somatic Cell Cloned Bovine Fetuses.
J. R. Hill, R. C. Burghardt, K. Jones, C. R. Long, C. R. Looney, T. Shin, T. E. Spencer, J. A. Thompson, Q. A. Winger, and M. E. Westhusin (2000)
Biol Reprod 63, 1787-1794
   Abstract »    Full Text »
In Vitro Development of Reconstructed Porcine Oocytes after Somatic Cell Nuclear Transfer.
D.-B. Koo, Y.-K. Kang, Y.-H. Choi, J. S. Park, S.-K. Han, I. Y. Park, S.-U. Kim, K.-K. Lee, D.-S. Son, W.-K. Chang, et al. (2000)
Biol Reprod 63, 986-992
   Abstract »    Full Text »
Advances in Livestock Nuclear Transfer.
B. Kühholzer and R. S. Prather (2000)
Experimental Biology and Medicine 224, 240-245
   Abstract »    Full Text »
Pig Cloning by Microinjection of Fetal Fibroblast Nuclei.
A. Onishi, M. Iwamoto, T. Akita, S. Mikawa, K. Takeda, T. Awata, H. Hanada, and A. C. F. Perry (2000)
Science 289, 1188-1190
   Abstract »    Full Text »
Increased Birefringence in the Meiotic Spindle Provides a New Marker for the Onset of Activation in Living Oocytes.
L. Liu, J. R. Trimarchi, R. Oldenbourg, and D. L. Keefe (2000)
Biol Reprod 63, 251-258
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Postnatal Growth and Behavioral Development of Mice Cloned from Adult Cumulus Cells.
K. L.K. Tamashiro, T. Wakayama, R. J. Blanchard, D. C. Blanchard, and R. Yanagimachi (2000)
Biol Reprod 63, 328-334
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Production of Male Cloned Mice from Fresh, Cultured, and Cryopreserved Immature Sertoli Cells.
A. Ogura, K. Inoue, N. Ogonuki, A. Noguchi, K. Takano, R. Nagano, O. Suzuki, J. Lee, F. Ishino, and J. Matsuda (2000)
Biol Reprod 62, 1579-1584
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Establishment of a Porcine Cell Line from In Vitro-Produced Blastocysts and Transfer of the Cells into Enucleated Oocytes.
K. Miyoshi, Y. Taguchi, Y. Sendai, H. Hoshi, and E. Sato (2000)
Biol Reprod 62, 1640-1646
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Alternative sources of gametes: reality or science fiction?.
M. C. Tsai, T. Takeuchi, J.M. Bedford, M. M. Reis, Z. Rosenwaks, and G. D. Palermo (2000)
Hum. Reprod. 15, 988-998
   Abstract »    Full Text »    PDF »
Chemically and mechanically induced membrane fusion: non-activating methods for nuclear transfer in mature human oocytes.
J. Tesarik, Z. P. Nagy, C. Mendoza, and E. Greco (2000)
Hum. Reprod. 15, 1149-1154
   Abstract »    Full Text »    PDF »
Development Rates of Male Bovine Nuclear Transfer Embryos Derived from Adult and Fetal Cells.
J. R. Hill, Q. A. Winger, C. R. Long, C. R. Looney, J. A. Thompson, and M. E. Westhusin (2000)
Biol Reprod 62, 1135-1140
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Extension of Cell Life-Span and Telomere Length in Animals Cloned from Senescent Somatic Cells.
R. P. Lanza, J. B. Cibelli, C. Blackwell, V. J. Cristofalo, M. K. Francis, G. M. Baerlocher, J. Mak, M. Schertzer, E. A. Chavez, N. Sawyer, et al. (2000)
Science 288, 665-669
   Abstract »    Full Text »
Analysis of Gene Transcription in Bovine Nuclear Transfer Embryos Reconstructed with Granulosa Cell Nuclei.
R. Daniels, V. Hall, and A.O. Trounson (2000)
Biol Reprod 63, 1034-1040
   Abstract »    Full Text »


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