Nature Biotechnology
20, 689 - 696 (2002)
Published online: 3 June 2002; | doi:10.1038/nbt703
Generation of histocompatible tissues using nuclear
transplantationRobert P. Lanza1, Ho Yun Chung2, James J. Yoo2, Peter J. Wettstein3, Catherine Blackwell1, Nancy Borson3, Erik Hofmeister3, Gunter Schuch2, Shay Soker2, Carlos T. Moraes4, Michael D. West1
& Anthony Atala21
Advanced Cell Technology,
Worcester, MA 01605. 2
Laboratory for Tissue Engineering and Cellular
Therapeutics, Children's Hospital and Harvard Medical School,
Boston, MA 02115. 3
Departments of Surgery and Immunology, Mayo
Clinic, Rochester, MN 55905. 4
Department of Neurology, University of Miami
School of Medicine, Miami, FL 33136.
Correspondence should be addressed to Anthony Atala Anthony.atala@TCH.Harvard.eduNuclear 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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