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Title: Glucose responsive insulin production from human embryonic germ (EG) cell derivatives

Abstract

Type 1 diabetes mellitus subjects millions to a daily burden of disease management, life threatening hypoglycemia and long-term complications such as retinopathy, nephropathy, heart disease, and stroke. Cell transplantation therapies providing a glucose-regulated supply of insulin have been implemented clinically, but are limited by safety, efficacy and supply considerations. Stem cells promise a plentiful and flexible source of cells for transplantation therapies. Here, we show that cells derived from human embryonic germ (EG) cells express markers of definitive endoderm, pancreatic and {beta}-cell development, glucose sensing, and production of mature insulin. These cells integrate functions necessary for glucose responsive regulation of preproinsulin mRNA and expression of insulin C-peptide in vitro. Following transplantation into mice, cells become insulin and C-peptide immunoreactive and produce plasma C-peptide in response to glucose. These findings suggest that EG cell derivatives may eventually serve as a source of insulin producing cells for the treatment of diabetes.

Authors:
 [1];  [2];  [2];  [2];  [2];  [3]
  1. Division of Endocrinology and Metabolism, Johns Hopkins University School of Medicine, 1830 E. Monument Street, Suite 333, Baltimore, MD 21287 (United States)
  2. Department of Gynecology and Obstetrics, Institute for Cell Engineering, Johns Hopkins University School of Medicine, 733 North Broadway, BRB 769, Baltimore, MD 21205 (United States)
  3. Department of Gynecology and Obstetrics, Institute for Cell Engineering, Johns Hopkins University School of Medicine, 733 North Broadway, BRB 769, Baltimore, MD 21205 (United States). E-mail: mshambl1@jhmi.edu
Publication Date:
OSTI Identifier:
20991333
Resource Type:
Journal Article
Resource Relation:
Journal Name: Biochemical and Biophysical Research Communications; Journal Volume: 356; Journal Issue: 3; Other Information: DOI: 10.1016/j.bbrc.2007.03.017; PII: S0006-291X(07)00474-3; Copyright (c) 2007 Elsevier Science B.V., Amsterdam, The Netherlands, All rights reserved; Country of input: International Atomic Energy Agency (IAEA)
Country of Publication:
United States
Language:
English
Subject:
60 APPLIED LIFE SCIENCES; CARDIOVASCULAR DISEASES; DIABETES MELLITUS; GENE REGULATION; GLUCOSE; IN VITRO; INSULIN; MICE; PANCREAS; PEPTIDES; STEM CELLS; THERAPY

Citation Formats

Clark, Gregory O., Yochem, Robert L., Axelman, Joyce, Sheets, Timothy P., Kaczorowski, David J., and Shamblott, Michael J. Glucose responsive insulin production from human embryonic germ (EG) cell derivatives. United States: N. p., 2007. Web. doi:10.1016/j.bbrc.2007.03.017.
Clark, Gregory O., Yochem, Robert L., Axelman, Joyce, Sheets, Timothy P., Kaczorowski, David J., & Shamblott, Michael J. Glucose responsive insulin production from human embryonic germ (EG) cell derivatives. United States. doi:10.1016/j.bbrc.2007.03.017.
Clark, Gregory O., Yochem, Robert L., Axelman, Joyce, Sheets, Timothy P., Kaczorowski, David J., and Shamblott, Michael J. Fri . "Glucose responsive insulin production from human embryonic germ (EG) cell derivatives". United States. doi:10.1016/j.bbrc.2007.03.017.
@article{osti_20991333,
title = {Glucose responsive insulin production from human embryonic germ (EG) cell derivatives},
author = {Clark, Gregory O. and Yochem, Robert L. and Axelman, Joyce and Sheets, Timothy P. and Kaczorowski, David J. and Shamblott, Michael J.},
abstractNote = {Type 1 diabetes mellitus subjects millions to a daily burden of disease management, life threatening hypoglycemia and long-term complications such as retinopathy, nephropathy, heart disease, and stroke. Cell transplantation therapies providing a glucose-regulated supply of insulin have been implemented clinically, but are limited by safety, efficacy and supply considerations. Stem cells promise a plentiful and flexible source of cells for transplantation therapies. Here, we show that cells derived from human embryonic germ (EG) cells express markers of definitive endoderm, pancreatic and {beta}-cell development, glucose sensing, and production of mature insulin. These cells integrate functions necessary for glucose responsive regulation of preproinsulin mRNA and expression of insulin C-peptide in vitro. Following transplantation into mice, cells become insulin and C-peptide immunoreactive and produce plasma C-peptide in response to glucose. These findings suggest that EG cell derivatives may eventually serve as a source of insulin producing cells for the treatment of diabetes.},
doi = {10.1016/j.bbrc.2007.03.017},
journal = {Biochemical and Biophysical Research Communications},
number = 3,
volume = 356,
place = {United States},
year = {Fri May 11 00:00:00 EDT 2007},
month = {Fri May 11 00:00:00 EDT 2007}
}
  • Research highlights: {yields} Human CD59 (hCD59) gene was introduced into porcine embryonic germ (EG) cells. {yields} hCD59-transgenic EG cells were resistant to hyperacute rejection in cytolytic assay. {yields} hCD59-transgenic pigs were produced by EG cell nuclear transfer. -- Abstract: This study was performed to produce transgenic pigs expressing the human complement regulatory protein CD59 (hCD59) using the nuclear transfer (NT) of embryonic germ (EG) cells, which are undifferentiated stem cells derived from primordial germ cells. Because EG cells can be cultured indefinitely in an undifferentiated state, they may provide an inexhaustible source of nuclear donor cells for NT to producemore » transgenic pigs. A total of 1980 NT embryos derived from hCD59-transgenic EG cells were transferred to ten recipients, resulting in the birth of fifteen piglets from three pregnancies. Among these offspring, ten were alive without overt health problems. Based on PCR analysis, all fifteen piglets were confirmed as hCD59 transgenic. The expression of the hCD59 transgene in the ten living piglets was verified by RT-PCR. Western analysis showed the expression of the hCD59 protein in four of the ten RT-PCR-positive piglets. These results demonstrate that hCD59-transgenic pigs could effectively be produced by EG cell NT and that such transgenic pigs may be used as organ donors in pig-to-human xenotransplantation.« less
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