FGF-signalling in the differentiation of mouse ES cells towards ...

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PrefaceThis thesis is based on experimental work performed in the Department of Stem Cell Biology(formerly the Department of Developmental Biology) at the Hagedorn Research Institute inGentofte – Denmark, from August 2006 to March 2010 (including an 8 month leave). Theproject supervisors were Berthe M. Willumsen, Professor at the Institute of Biology, Faculty ofScience, University of Copenhagen – Denmark, and Palle Serup, Ph.D., Scientific Director atthe Hagedorn Research Institute, Gentofte – Denmark. The work was supported by grants fromthe Juvenile Diabetes Research Foundation and the Beta Cell Biology Consortium, NationalInstitutes of Health – USA.This thesis is submitted in order to meet the requirements for obtaining the degree ofphilosophiæ doctor (Ph.D.) at the Faculty of Science, University of Copenhagen – Denmark.The thesis is based on two papers:Paper I:‘A late requirement for Wnt and FGF signaling during Activin-induced formation of foregutendoderm from mouse embryonic stem cells’Published in Developmental Biology, 2009; 330, pp. 286-304.Mattias Hansson, Dorthe R- Olesen, Janny M.L. Peterslund, Nina Engberg, Morten Kahn,Maria Winzi, Tino Klein, Poul Maddox-Hyttel and Palle SerupPaper II:‘FGFR(IIIc)-activation induces mesendoderm but is dispensable for definitive endodermformation in mouse embryonic stem cells’Manuscript to be submitted.Janny Marie L. Peterslund and Palle SerupIn addition, this thesis contains a general introduction to early mouse embryonic developmentand pancreas formation, directed differentiation of mES cells towards DE and pancreatic β-likecells, and FGF-signalling in relation to both development and differentiation. The reader willalso find a short chapter expanding on the patterning of definitive endoderm described in paperI and a general discussion of my work in relation to the research field as a whole.Janny Marie Landegent PeterslundMarch 2010III
Page 1: PhD thesisCand.scient. Janny Marie
Page 8 and 9: VIII
Page 11 and 12: AbbreviationsAP1 activator protein
Page 13 and 14: 1. General introductionDiabetes mel
Page 15 and 16: SOX2 and OCT4. Later, the PE and ep
Page 17 and 18: (CIP) move rostrally (arrows in Fig
Page 19 and 20: signalling. These dynamic signallin
Page 21: Figure 1-4: Key signalling pathways
Page 26 and 27: alternative splicing of exons 8 and
Page 29 and 30: 3. Paper IA late requirement for Wn
Page 31 and 32: M. Hansson et al. / Developmental B
Page 49 and 50: Supplementary data, Paper I‘A lat
Page 51 and 52: Figure S3Figure S3: ES cell progeny
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Figure S547
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4. Pxd1 inductionBackgroundThe work
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Figure 4-2: WNT3a and FGF4 in Step
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Figure 4-5: Pdx1 cannot be detected
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DE in medium containing activin onl
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FGFR(IIIc)-activation induces mesen
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IntroductionBased on knowledge obta
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For analysis of cells stained with
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they were divided into three catego
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and 9 being the most effective ones
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Most of the remaining cells are Sox
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used in this paper. Modified from (
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Gadue, P., T.L. Huber, P.J. Paddiso
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Ying, Q.L., J. Nichols, I. Chambers
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Figure 2: Activation of FGFRc-isofo
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Figure 4: FGFs activating FGFRc-iso
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Figure S2: A few areas of SOX7 + ce
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The definitive problem of endodermW
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involved in WNT-signalling and main
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2008). Lineage tracing of these cel
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7. ReferencesAllen, B.L. and A.C. R
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Gadue, P., T.L. Huber, P.J. Paddiso
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Madsen, O.D. and P. Serup. 2006. To
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Rodaway, A. and R. Patient. 2001. M
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Yamanaka, S. 2009. A fresh look at
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