Isolation and culture of endothelial cells from embryonic rat yolk sac

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dc.contributor.authorUlger, Harun
dc.contributor.authorKarabulut, Ahmet K.
dc.contributor.authorPratten, Margaret K.
dc.date.accessioned2020-03-26T19:41:58Z
dc.date.available2020-03-26T19:41:58Z
dc.date.issued2017
dc.departmentSelçuk Üniversitesien_US
dc.description.abstractYolk sac blood islands are the first morphologic evidence of hematopoietic development during mammalian embryogenesis, and visseral yolk sac mesoderm gives rise to the first embryonic blood cells within a rich endothelial network. Present study reports the isolation and culture of endothelial cells from 11.5 days old embryonic rat yolk sac. The embryos were dissected from 11.5 days pregnant Wistar rat (Rattus norvegicus) and the external yolk sac membrane and embryos were removed under aseptic condition. After washing three times with Calcium-Magnesium free Hank's balanced salt solution (CMF-HBSS), the tissue was minced, and fragments were incubated in CMF-HBSS containing 2mg/ml Trypsin, 100mg/ml collagenase I and 40mg/ml DNAse at 37 degrees C until the tissue was completely dispersed. The digestion effect was then neutralized by fetal bovine serum at 1:3 (v/v). The cell suspension was centrifuged at 1000 rpm for 10 min., the supernatants were discarded and the cell pellets resuspended in Dulbecco modified Eagle medium containing 15% fetal bovine serum, 1.25mg/ml amphotericin B, 25mg/ml gentamycin sulphate and 100mg/ml endothelial cell growth supplement. The resuspended cells were plated in two diverse 25cm(2) culture flasks for overnight differential adherence at 37 degrees C. The non-adherent cells were removed by gentle aspiration and adherent cells refed with fresh medium. The cells were transferred using 1ml of 0.2% Trypsin when cultures reached near-confluence. The cultured yolk sac endothelial cells had characteristic cobblestone appearence and positive immunofluorescent staining for von Willebrand Factor (vWF). Weibel-Palade bodies, the major ultrastructural marker for endothelium, were also detected in cultured cells by electron microscopy.en_US
dc.identifier.doi10.24190/ISSN2564-615X/2017/02.06en_US
dc.identifier.endpage154en_US
dc.identifier.issn2564-615Xen_US
dc.identifier.issue2en_US
dc.identifier.pmid#YOKen_US
dc.identifier.startpage149en_US
dc.identifier.urihttps://dx.doi.org/10.24190/ISSN2564-615X/2017/02.06
dc.identifier.urihttps://hdl.handle.net/20.500.12395/35243
dc.identifier.volume1en_US
dc.identifier.wosWOS:000467955700006en_US
dc.identifier.wosqualityN/Aen_US
dc.indekslendigikaynakWeb of Scienceen_US
dc.language.isoenen_US
dc.publisherSCIENDOen_US
dc.relation.ispartofEUROBIOTECH JOURNALen_US
dc.relation.publicationcategoryMakale - Uluslararası Hakemli Dergi - Kurum Öğretim Elemanıen_US
dc.rightsinfo:eu-repo/semantics/openAccessen_US
dc.selcuk20240510_oaigen_US
dc.titleIsolation and culture of endothelial cells from embryonic rat yolk sacen_US
dc.typeArticleen_US

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