Human oocytes reprogram adult somatic nuclei of a type 1 diabetic to diploid pluripotent stem cells.
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.
1] The New York Stem Cell Foundation Research Institute, New York, New York 10032, USA [2].
Mitsutoshi
M
Yamada
1] The New York Stem Cell Foundation Research Institute, New York, New York 10032, USA [2].
Bjarki
B
Johannesson
Stem Cell Unit, Department of Genetics, Silberman Institute of Life Sciences, The Hebrew University of Jerusalem, Jerusalem 91904, Israel.
Ido
I
Sagi
Naomi Berrie Diabetes Center, Department of Pediatrics, College of Physicians and Surgeons, Columbia University, New York, New York 10032, USA.
Lisa Cole
LC
Burnett
1] Center for Women's Reproductive Care, College of Physicians and Surgeons, Columbia University, New York 10019, USA [2] Department of Obstetrics and Gynecology, College of Physicians and Surgeons, Columbia University, New York 10032, USA.
Daniel H
DH
Kort
1] Center for Women's Reproductive Care, College of Physicians and Surgeons, Columbia University, New York 10019, USA [2] Department of Obstetrics and Gynecology, College of Physicians and Surgeons, Columbia University, New York 10032, USA.
Robert W
RW
Prosser
The New York Stem Cell Foundation Research Institute, New York, New York 10032, USA.
Daniel
D
Paull
The New York Stem Cell Foundation Research Institute, New York, New York 10032, USA.
Michael W
MW
Nestor
Naomi Berrie Diabetes Center, Department of Pediatrics, College of Physicians and Surgeons, Columbia University, New York, New York 10032, USA.
Matthew
M
Freeby
Naomi Berrie Diabetes Center, Department of Pediatrics, College of Physicians and Surgeons, Columbia University, New York, New York 10032, USA.
Ellen
E
Greenberg
Naomi Berrie Diabetes Center, Department of Pediatrics, College of Physicians and Surgeons, Columbia University, New York, New York 10032, USA.
Robin S
RS
Goland
Naomi Berrie Diabetes Center, Department of Pediatrics, College of Physicians and Surgeons, Columbia University, New York, New York 10032, USA.
Rudolph L
RL
Leibel
The New York Stem Cell Foundation Research Institute, New York, New York 10032, USA.
Susan L
SL
Solomon
Stem Cell Unit, Department of Genetics, Silberman Institute of Life Sciences, The Hebrew University of Jerusalem, Jerusalem 91904, Israel.
Nissim
N
Benvenisty
1] Center for Women's Reproductive Care, College of Physicians and Surgeons, Columbia University, New York 10019, USA [2] Department of Obstetrics and Gynecology, College of Physicians and Surgeons, Columbia University, New York 10032, USA.
Mark V
MV
Sauer
The New York Stem Cell Foundation Research Institute, New York, New York 10032, USA.
Dieter
D
Egli
D000328:Adult; D001755:Blastocyst / Q000187:drug effects; D002459:Cell Fusion; D002467:Cell Nucleus / Q000235:genetics*; D065150:Cellular Reprogramming; D033481:Chromosomes, Mammalian / Q000378:metabolism; D003922:Diabetes Mellitus, Type 1 / Q000235:genetics* / Q000473:pathology*; D004171:Diploidy; D005260:Female; D056572:Histone Deacetylase Inhibitors / Q000494:pharmacology; D006801:Humans; D007231:Infant, Newborn; D008677:Metaphase; D009865:Oocytes / Q000166:cytology* / Q000378:metabolism; D009866:Oogenesis; D039904:Pluripotent Stem Cells / Q000166:cytology* / Q000378:metabolism / Q000473:pathology; D029082:Sendai virus; D008941:Spindle Apparatus / Q000378:metabolism
D056572:Histone Deacetylase Inhibitors