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dc.contributor.authorAldrich, Jessica
dc.contributor.authorLong, David S.
dc.date.accessioned2019-05-30T21:03:52Z
dc.date.available2019-05-30T21:03:52Z
dc.date.issued2019-05-03
dc.identifier.citationAldrich, Jessica; Long, David S. 2019. In situ fixation and subsequent collection of cultured endothelial cells in a shear flow. MethodsX, vol. 6:pp 1164-1173en_US
dc.identifier.issn2215-0161
dc.identifier.urihttps://doi.org/10.1016/j.mex.2019.05.001
dc.identifier.urihttp://hdl.handle.net/10057/16323
dc.description© 2019 The Authors. This article is available under the terms of the Creative Commons Attribution License (CC BY).en_US
dc.description.abstractIn situ fixation of adherent cells is a necessary process for downstream assays. Current methods to dissociate adherent endothelial cells require the use of a cell scraper that may introduce variability in nuclear morphology. Also, a cell scraper is not an option for experiments using sealed flow chambers. HMEC-1 cells were sheared at 5 dyn/cm 2 for 24 h and then fixed in situ, quenched, and dissociated at the same shear rate. Analysis revealed no statistically significant change in nuclear shape between the steps of fixation and dissociation. This method outlines an alternative for the dissociation of adherent sheared endothelial cells after being fixed in situ in a micro-scale channel without causing a change in the nuclear morphology. • This method can be used with any commercially available, or custom-made, flow chamber and flow system. • Allows for downstream experimentation with adherent cells fixed in situ, such as Hi-C analysis, without impacting nuclear morphology or chromatin organization. • Cells are cultured, fixed, and dissociated at the same shear rate. Using the same shear rate for each step yields results that are not influenced by variable forces.en_US
dc.description.sponsorshipCollege of Engineering at Wichita State University (D.S.L.). J.L.A. received partial support from the Kansas Idea Network of Biomedical Research Excellence , (Institutional Development Award (IDeA) from the National Institute of General Medical Sciences of the National Institutes of Health under grant number P20GM103418 ).en_US
dc.language.isoen_USen_US
dc.publisherElsevier B.V.en_US
dc.relation.ispartofseriesMethodsX;v.6
dc.subjectAdherent cellsen_US
dc.subjectCell dissociationen_US
dc.subjectChromatin conformation captureen_US
dc.subjectEndothelial cellsen_US
dc.subjectEndotheliumen_US
dc.subjectHi-Cen_US
dc.subjectIn situ fixationen_US
dc.subjectIn situ fixation and subsequent collection of cultured endothelial cells in a shear flowen_US
dc.subjectMechanogenomicsen_US
dc.titleIn situ fixation and subsequent collection of cultured endothelial cells in a shear flowen_US
dc.typeArticleen_US
dc.rights.holder© 2019 The Authors. Published by Elsevier B.V.en_US


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