Human oocytes reprogram adult somatic nuclei of a type 1 diabetic to diploid pluripotent stem cells.

Authors:
Mitsutoshi Yamada
Mitsutoshi Yamada
Keio University School of Medicine
Japan
Bjarki Johannesson
Bjarki Johannesson
University of Heidelberg
Germany
Ido Sagi
Ido Sagi
Silberman Institute of Life Sciences
Israel
Lisa Cole Burnett
Lisa Cole Burnett
Naomi Berrie Diabetes Center
New York | United States
Daniel H Kort
Daniel H Kort
Columbia University College of Physicians and Surgeons
New York | United States
Robert W Prosser
Robert W Prosser
College of Physicians and Surgeons of Columbia University
United States
Daniel Paull
Daniel Paull
Johns Hopkins University
United States
Michael W Nestor
Michael W Nestor
University of Maryland School of Medicine
United States

Nature 2014 Jun 28;510(7506):533-6. Epub 2014 Apr 28.

The New York Stem Cell Foundation Research Institute, New York, New York 10032, USA.

The transfer of somatic cell nuclei into oocytes can give rise to pluripotent stem cells that are consistently equivalent to embryonic stem cells, holding promise for autologous cell replacement therapy. Although methods to induce pluripotent stem cells from somatic cells by transcription factors are widely used in basic research, numerous differences between induced pluripotent stem cells and embryonic stem cells have been reported, potentially affecting their clinical use. Because of the therapeutic potential of diploid embryonic stem-cell lines derived from adult cells of diseased human subjects, we have systematically investigated the parameters affecting efficiency of blastocyst development and stem-cell derivation. Here we show that improvements to the oocyte activation protocol, including the use of both kinase and translation inhibitors, and cell culture in the presence of histone deacetylase inhibitors, promote development to the blastocyst stage. Developmental efficiency varied between oocyte donors, and was inversely related to the number of days of hormonal stimulation required for oocyte maturation, whereas the daily dose of gonadotropin or the total number of metaphase II oocytes retrieved did not affect developmental outcome. Because the use of concentrated Sendai virus for cell fusion induced an increase in intracellular calcium concentration, causing premature oocyte activation, we used diluted Sendai virus in calcium-free medium. Using this modified nuclear transfer protocol, we derived diploid pluripotent stem-cell lines from somatic cells of a newborn and, for the first time, an adult, a female with type 1 diabetes.

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June 2014
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