{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2023,12,24]],"date-time":"2023-12-24T23:50:49Z","timestamp":1703461849211},"reference-count":33,"publisher":"Wiley","issue":"5","license":[{"start":{"date-parts":[[2009,5,7]],"date-time":"2009-05-07T00:00:00Z","timestamp":1241654400000},"content-version":"vor","delay-in-days":0,"URL":"http:\/\/onlinelibrary.wiley.com\/termsAndConditions#vor"}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["Diabetes Metabolism Res"],"published-print":{"date-parts":[[2009,7]]},"abstract":"<jats:title>Abstract<\/jats:title><jats:sec><jats:title>Background<\/jats:title><jats:p>Embryonic stem (ES) cells have the potential to produce unlimited numbers of surrogate insulin\u2010producing cells for cell replacement therapy of type 1 diabetes mellitus. The impact of the <jats:italic>in vivo<\/jats:italic> environment on mouse ES cell differentiation towards insulin\u2010producing cells was analysed morphologically after implantation.<\/jats:p><\/jats:sec><jats:sec><jats:title>Methods<\/jats:title><jats:p>ES cells differentiated <jats:italic>in vitro<\/jats:italic> into insulin\u2010producing cells according to the Lumelsky protocol or a new four\u2010stage differentiation protocol were analysed morphologically before and after implantation for gene expression by <jats:italic>in situ<\/jats:italic> reverse transcription polymerase chain reaction and protein expression by immunohistochemistry and ultrastructural analysis.<\/jats:p><\/jats:sec><jats:sec><jats:title>Results<\/jats:title><jats:p>In comparison with nestin positive ES cells developed according to the reference protocol, the number of ES cells differentiated with the four\u2010stage protocol increased under <jats:italic>in vivo<\/jats:italic> conditions upon morphological analysis. The cells exhibited, in comparison to the <jats:italic>in vitro<\/jats:italic> situation, increased gene and protein expression of Pdx1, insulin, islet amyloid polypeptide (IAPP), the GLUT2 glucose transporter and glucokinase, which are functional markers for glucose\u2010induced insulin secretion of pancreatic beta cells. Renal sub\u2010capsular implantation of ES cells with a higher degree of differentiation achieved by <jats:italic>in vitro<\/jats:italic> differentiation with a four\u2010stage protocol enabled further significant maturation for the beta\u2010cell\u2010specific markers, insulin and the co\u2010stored IAPP as well as the glucose recognition structures. In contrast, further <jats:italic>in vivo<\/jats:italic> differentiation was not achieved with cells differentiated <jats:italic>in vitro<\/jats:italic> by the reference protocol.<\/jats:p><\/jats:sec><jats:sec><jats:title>Conclusions<\/jats:title><jats:p>A sufficient degree of <jats:italic>in vitro<\/jats:italic> differentiation is an essential prerequisite for further substantial maturation in a beta\u2010cell\u2010specific way <jats:italic>in vivo<\/jats:italic>, supported by cell\u2010cell contacts and vascularisation. Copyright \u00a9 2009 John Wiley &amp; Sons, Ltd.<\/jats:p><\/jats:sec>","DOI":"10.1002\/dmrr.965","type":"journal-article","created":{"date-parts":[[2009,5,7]],"date-time":"2009-05-07T14:49:07Z","timestamp":1241707747000},"page":"464-476","source":"Crossref","is-referenced-by-count":15,"title":["Changes in gene expression and morphology of mouse embryonic stem cells on differentiation into insulin\u2010producing cells <i>in vitro<\/i> and <i>in vivo<\/i>"],"prefix":"10.1002","volume":"25","author":[{"given":"Ortwin","family":"Naujok","sequence":"first","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Flavio","family":"Francini","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Sally","family":"Picton","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Clifford 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