Please use this identifier to cite or link to this item: https://doi.org/10.1007/s00125-004-1458-8
Title: In vitro directed differentation of mouse embryonic stem cells into insulin-producing cells
Authors: León-Quinto, T.
Jones, J.
Skoudy, A.
Burcin, M.
Soria, B. 
Keywords: Diabetes
Differentiation
Embryonic stem cells
Exogenous agents
Insulin-producing cells
Islet progenitors
Issue Date: Aug-2004
Source: León-Quinto, T., Jones, J., Skoudy, A., Burcin, M., Soria, B. (2004-08). In vitro directed differentation of mouse embryonic stem cells into insulin-producing cells. Diabetologia 47 (8) : 1442-1451. ScholarBank@NUS Repository. https://doi.org/10.1007/s00125-004-1458-8
Abstract: Aims/hypothesis. We recently demonstrated that insulin-producing cells derived from embryonic stem cells normalise hyperglycaemia in transplanted diabetic mice. The differentiation and selection procedure, however, was successful in less than 5% of the assays performed. Thus, to improve its effectiveness, new strategies have been developed, which increase the number of islet cells or islet progenitors. Methods. Mouse embryonic stem cells transfected with a plasmid containing the Nkx6.1 promoter gene followed by a neomycin-resistance gene, were cultured with factors known to participate in endocrine pancreatic development and factors that modulate signalling pathways involved in these processes. Neomycin was used to select the Nkx6.1-positive cells, which also express insulin. The transfected cells were differentiated using several exogenous agents, followed by selection of Nkx6.1-positive cells. The resulting cells were analysed for pancreatic gene and protein expression by immunocytochemistry, RT-PCR and radioimmunoassay. Also, proliferation assays were performed, as well as transplantation to streptozotocin-induced diabetic mice. Results. The protocols yielded cell cultures with approximately 20% of cells co-expressing insulin and Pdx-1. Cell trapping selection yielded an almost pure population of insulin-positive cells, which expressed the beta cell genes/proteins Pdx-1, Nkx6.1, insulin, glucokinase, GLUT-2 and Sur-1. Subsequent transplantation to streptozotocin-induced diabetic mice normalised their glycaemia during the time period of experimentation, proving the efficiency of the protocols. Conclusions/interpretation. These methods were both highly efficient and very reproducible, resulting in a new strategy to obtain insulin-containing cells from stem cells with a near 100% success rate, while actively promoting the maturation of the exocytotic machinery. © Springer-Verlag 2004.
Source Title: Diabetologia
URI: http://scholarbank.nus.edu.sg/handle/10635/108113
ISSN: 0012186X
DOI: 10.1007/s00125-004-1458-8
Appears in Collections:Staff Publications

Show full item record
Files in This Item:
There are no files associated with this item.

SCOPUSTM   
Citations

150
checked on Feb 13, 2018

WEB OF SCIENCETM
Citations

122
checked on Jan 10, 2018

Page view(s)

23
checked on Feb 18, 2018

Google ScholarTM

Check

Altmetric


Items in DSpace are protected by copyright, with all rights reserved, unless otherwise indicated.