caa a22 4500 445292601 CHVBK 20180317142533.0 cr unu---uuuuu 170323e20100701xx s 000 0 eng 10.1007/s11626-010-9313-3 doi (NATIONALLICENCE)springer-10.1007/s11626-010-9313-3 Establishment of heparan sulphate deficient primary endothelial cells from EXT-1flox/flox mouse lungs and sprouting aortas [Elektronische Daten] [Paulina Kucharzewska, Johanna Welch, Josefin Birgersson, Mattias Belting] Angiogenesis is a hallmark of expanding tissue e.g. during embryogenesis and wound healing in physiology as well as in diseases such as cancer and atherosclerosis. Key steps of the angiogenic process involve growth factor-mediated stimulation of endothelial cell sprouting and tube formation. Heparan sulphate proteoglycans (HSPGs) have been implicated as important co-receptors of several pro-angiogenic proteins. The importance of HSPGs in physiology was underscored by the finding that knockout of the gene encoding HS polymerase, EXT-1, resulted in early embryonic lethality. Here, we describe the establishment of HS-deficient endothelial cells from sprouting aortas as well as from the lungs of EXT-1flox/flox mice. Recombination of the loxP-flanked EXT-1 locus by Cre-expressing adenovirus was demonstrated at the mRNA level. Moreover, depletion of HS polysaccharides was verified by flow cytometry and fluorescence microscopy methodology using phage display-derived anti-HS antibodies. In summary, we provide a genetic model to unravel the functional role of HSPGs specifically in primary endothelial cells during early steps of angiogenesis. Our studies are applicable to most loxP-based transgenic mouse strains, and may thus be of general importance in the angiogenesis field. The Society for In Vitro Biology, 2010 Angiogenesis nationallicence EXT-1 nationallicence Endothelial cells nationallicence Heparan sulphate proteoglycans nationallicence Adenoviral vector nationallicence Kucharzewska Paulina Department of Oncology, Clinical Sciences, Lund University and Lund University Hospital, Barngatan 2B, SE-221 85, Lund, Sweden aut Welch Johanna Department of Oncology, Clinical Sciences, Lund University and Lund University Hospital, Barngatan 2B, SE-221 85, Lund, Sweden aut Birgersson Josefin Department of Oncology, Clinical Sciences, Lund University and Lund University Hospital, Barngatan 2B, SE-221 85, Lund, Sweden aut Belting Mattias Department of Oncology, Clinical Sciences, Lund University and Lund University Hospital, Barngatan 2B, SE-221 85, Lund, Sweden aut In Vitro Cellular & Developmental Biology - Animal Springer-Verlag 46/7(2010-07-01), 577-584 1071-2690 46:7<577 2010 46 11626 https://doi.org/10.1007/s11626-010-9313-3 text/html Onlinezugriff via DOI 1 research-article jats NATIONALLICENCE 856 40 https://doi.org/10.1007/s11626-010-9313-3 text/html Onlinezugriff via DOI NATIONALLICENCE 700 1- Kucharzewska Paulina Department of Oncology, Clinical Sciences, Lund University and Lund University Hospital, Barngatan 2B, SE-221 85, Lund, Sweden aut NATIONALLICENCE 700 1- Welch Johanna Department of Oncology, Clinical Sciences, Lund University and Lund University Hospital, Barngatan 2B, SE-221 85, Lund, Sweden aut NATIONALLICENCE 700 1- Birgersson Josefin Department of Oncology, Clinical Sciences, Lund University and Lund University Hospital, Barngatan 2B, SE-221 85, Lund, Sweden aut NATIONALLICENCE 700 1- Belting Mattias Department of Oncology, Clinical Sciences, Lund University and Lund University Hospital, Barngatan 2B, SE-221 85, Lund, Sweden aut NATIONALLICENCE 773 0- In Vitro Cellular & Developmental Biology - Animal Springer-Verlag 46/7(2010-07-01), 577-584 1071-2690 46:7<577 2010 46 11626 Metadata rights reserved Springer special CC-BY-NC licence nationallicence BK010053 XK010053 XK010000 NATIONALLICENCE NATIONALLICENCE NL-springer