Producing primate embryonic stem cells by somatic cell nuclear transfer

• Published in: Nature 2007-11, Vol.450 (7169), p.497-502
• Main Authors: Mitalipov, S. M, Byrne, J. A, Pedersen, D. A, Clepper, L. L, Nelson, M, Sanger, W. G, Gokhale, S, Wolf, D. P
• Format: Article
• Language: English
• Subjects: Anesthesia. Intensive care medicine. Transfusions. Cell therapy and gene therapy; Animals; Applied cell therapy and gene therapy; Base Sequence; Biological and medical sciences; Biomedical research; Cell Differentiation; Cell morphology; Cells, Cultured; Cellular biology; Cloning; DNA, Mitochondrial - genetics; Embryonic growth stage; Embryonic Stem Cells - cytology; Embryonic Stem Cells - immunology; Embryonic Stem Cells - metabolism; Embryos; Female; Fibroblasts; Gene Expression Profiling; Humanities and Social Sciences; Humans; Immune system; Immunology; Macaca mulatta; Macaca mulatta - genetics; Macaca mulatta - metabolism; Male; Medical sciences; Mice; Microsatellite Repeats - genetics; Mitochondrial DNA; multidisciplinary; Nuclear Transfer Techniques; Organ Specificity; Pluripotent Stem Cells - cytology; Pluripotent Stem Cells - metabolism; Primates; Science; Science (multidisciplinary); Stem cells; Transcription, Genetic; Transfusions. Complications. Transfusion reactions. Cell and gene therapy
• ISSN: 0028-0836; 1476-4687; 1476-4679
• DOI: 10.1038/nature06357

Derivation of embryonic stem (ES) cells genetically identical to a patient by somatic cell nuclear transfer (SCNT) holds the potential to cure or alleviate the symptoms of many degenerative diseases while circumventing concerns regarding rejection by the host immune system. However, the concept has only been achieved in the mouse, whereas inefficient reprogramming and poor embryonic development characterizes the results obtained in primates. Here, we used a modified SCNT approach to produce rhesus macaque blastocysts from adult skin fibroblasts, and successfully isolated two ES cell lines from these embryos. DNA analysis confirmed that nuclear DNA was identical to donor somatic cells and that mitochondrial DNA originated from oocytes. Both cell lines exhibited normal ES cell morphology, expressed key stem-cell markers, were transcriptionally similar to control ES cells and differentiated into multiple cell types in vitro and in vivo . Our results represent successful nuclear reprogramming of adult somatic cells into pluripotent ES cells and demonstrate proof-of-concept for therapeutic cloning in primates. Embryonic stem cells Techniques for reprogramming adult cells are much sought after for creating cells to genetically match those of patients. Such embryonic stem cells might be used therapeutically without immune rejection. One reprogramming method is somatic cell nuclear transfer, where an adult cell nucleus is inserted into an oocyte that has its own nucleus removed. An early (blastocyst) stage embryo is induced to form, from which embryonic stem cells are teased out and cultured. Previously, this was possible only in mice. Now Byrne et al . have succeeded with the technique using primate adult fibroblasts as the start point. They generated two embryonic stem cell lines from 304 oocytes taken from 14 rhesus monkeys. This success suggests that this approach might be suitable for generating patient-derived embryonic stem cells. A validation paper accompanies this package on the Nature web site. The colorized cover image shows an egg just prior to enucleation. The egg is held by a holding pipette and to the right, a sharp enucleation pipette points towards the nucleus. The reprogramming of rhesus macaque adult fibroblasts into embryonic stem cells using somatic cell nuclear transfer is demonstrated. Until now, creating embryonic stem cells in this way has only been successful in mice. This success with primates suggests that this approach could work for g