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Generation of histocompatible tissues using nuclear transplantation

Nature Biotechnology volume 20, pages 689696 (2002) | Download Citation

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Abstract

Nuclear transplantation (therapeutic cloning) could theoretically provide a limitless source of cells for regenerative therapy. Although the cloned cells would carry the nuclear genome of the patient, the presence of mitochondria inherited from the recipient oocyte raises questions about the histocompatibility of the resulting cells. In this study, we created bioengineered tissues from cardiac, skeletal muscle, and renal cells cloned from adult bovine fibroblasts. Long-term viability was demonstrated after transplantation of the grafts into the nuclear donor animals. Reverse transcription-PCR (RT-PCR) and western blot analysis confirmed that the cloned tissues expressed tissue-specific mRNA and proteins while expressing a different mitochondrial DNA (mtDNA) haplotype. In addition to creating skeletal muscle and cardiac “patches”, nuclear transplantation was used to generate functioning renal units that produced urinelike fluid and demonstrated unidirectional secretion and concentration of urea nitrogen and creatinine. Examination of the explanted renal devices revealed formation of organized glomeruli- and tubule-like structures. Delayed-type hypersensitivity (DTH) testing in vivo and Elispot analysis in vitro suggested that there was no rejection response to the cloned renal cells. The ability to generate histocompatible cells using cloning techniques addresses one of the major challenges in transplantation medicine.

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Acknowledgements

We thank Jose B. Cibelli, Frederick F. Hess (DVM), R. T. Duby, and the Department of Veterinary Sciences, University of Massachusetts, Amherst. We also thank Kyung-Ha Kang (Brigham & Women's Hospital), Wendy Nevala (Mayo Clinic), and Maria P. Bayona-Bafaluy (University of Miami) for their help with the histologic, Elispot, and molecular analyses. This research was supported in part by National Institutes of Health grants AI-16052 (to P.J.W.) and RO1DK57260 (to A.A.).

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Affiliations

  1. Advanced Cell Technology, Worcester, MA 01605.

    • Robert P. Lanza
    • , Catherine Blackwell
    •  & Michael D. West
  2. Laboratory for Tissue Engineering and Cellular Therapeutics, Children's Hospital and Harvard Medical School, Boston, MA 02115.

    • Ho Yun Chung
    • , James J. Yoo
    • , Gunter Schuch
    • , Shay Soker
    •  & Anthony Atala
  3. Departments of Surgery and Immunology, Mayo Clinic, Rochester, MN 55905.

    • Peter J. Wettstein
    • , Nancy Borson
    •  & Erik Hofmeister
  4. Department of Neurology, University of Miami School of Medicine, Miami, FL 33136.

    • Carlos T. Moraes

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Competing interests

R.P.L., C.B., and M.D.W. are employed by Advanced Cell Technology, which is pursuing therapeutic cloning.

Corresponding author

Correspondence to Anthony Atala.

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DOI

https://doi.org/10.1038/nbt703

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