The role of the adapter protein SHB in embryonic stem cell differentiation into the pancreatic beta-cell and endothelial lineages.
Embryonic stem (ES) cells represent an attractive tool not only for the study of the development of various cell types but also as a potential source of cells for transplantation. Previous studies suggested a role of the signal transduction protein SRC homology 2(SH2) protein of Beta-cells (SHB) for the development of both pancreatic 3-cells and blood vessels. SHB is an SH2 domain-containing adapter protein involved in the generation of signaling complexes in response to activation of a variety of receptors, several of which have been implicated in developmental processes. Moreover, microarray analysis of ES cells expressing mutant SHB has revealed decreased expression of several genes of developmental importance. Here, we present protocols that may be used for transfection of mouse ES cells and to study the differentiation of ES cell-derived embryoid bodies (EBs) into the pancreatic Beta-cell lineage as well as into vascular structures with special reference to the effect of SHB. Moreover, we also provide a protocol that may be used for enrichment by fluorescence-activated cell sorting of specific cell lineages in EBs.
Saldeen
Johan
J
Department of Medical Cell Biology, Uppsala University, Sweden.
Agren
Nina
N
Akerblom
Björn
B
Lu
Lingge
L
Adem
Bashir
B
Sedek
Lukasz
L
Kriz
Vitezslav
V
D000818:Animals; D018929:Cell Culture Techniques; D002454:Cell Differentiation / Q000502:physiology*; D019070:Cell Lineage; D004622:Embryo, Mammalian / Q000166:cytology; D042783:Endothelial Cells / Q000166:cytology / Q000502:physiology*; D005347:Fibroblasts / Q000166:cytology / Q000378:metabolism; D049452:Green Fluorescent Proteins / Q000378:metabolism; D050417:Insulin-Secreting Cells / Q000166:cytology / Q000502:physiology*; D051379:Mice; D011518:Proto-Oncogene Proteins / Q000235:genetics / Q000378:metabolism*; D020133:Reverse Transcriptase Polymerase Chain Reaction; D013234:Stem Cells / Q000166:cytology / Q000502:physiology*
D011518:Proto-Oncogene Proteins; C085634:Shb protein, mouse; D049452:Green Fluorescent Proteins