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Nuclei of Nonviable Ovine Somatic Cells Develop into Lambs after Nuclear Transplantation

^a Dipartimento di Strutture, Funzioni, Patologie e Biotecnologie, Università di Teramo, Italy ^b Department of Gene Expression & Development, The Roslin Institute, United Kingdom ^c Research Institute of Animal Science, Praha, Czech Republic ^d Istituto Zootecnico Caseario, 07040 Olmedo, Italy ^e Institute of Molecular Genetics, CNRS, UMR-5535, Montpellier, France ^f Laboratory of Protein Function, The Babraham Institute, Babraham, Cambridge, United Kingdom ^g Department of Animal Reproduction, University of Agriculture, Krakòw, Poland

Here we report on the successful reprogramming of nuclei from somatic cells rendered nonviable by heat treatment. Granulosa cells from adult sheep were heated to nonphysiological temperatures (55°C or 75°C) before their nuclei were injected into enucleated metaphase II oocytes. Reprogramming was demonstrated by the capacity of the reconstructed embryos to develop to the blastocyst stage in vitro and into fetuses and viable offspring in suitable foster mothers. To our knowledge, this is the first report of cloned mammalian offspring originating from nonviable cells. In addition, our experiments show that heat-treating donor nuclei destabilizes higher-order features of chromatin (but leaves intact its nucleosomal organization) and results in a high proportion of reconstructed embryos developing to the blastocyst stage and beyond.

First decision: 25 October 2001.

¹ Part of this work was supported by the Royal Society (to R.F., G.P., and P.L.). J.F. Jr. acknowledges the support from GACR 524/96/K162 and Mze NAZV EP 0960006200. Part of this work was covered by Es. Fin. 2001 (MIUR).

² Correspondence: Pasqualino Loi, Dipartimento di Strutture, Funzioni, Patologie e Biotecnologie, Facoltà di Medicina Veterinaria, Piazza Aldo Moro 45, 64100 Teramo, Italy. FAX: 0861 558819; loi{at}ifv.vet.unite.it

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