{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,9,4]],"date-time":"2026-09-04T09:04:20Z","timestamp":1788512660407,"version":"build-2803163510"},"reference-count":30,"publisher":"Oxford University Press (OUP)","issue":"3","license":[{"start":{"date-parts":[[2021,12,17]],"date-time":"2021-12-17T00:00:00Z","timestamp":1639699200000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/academic.oup.com\/journals\/pages\/open_access\/funder_policies\/chorus\/standard_publication_model"}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":[],"published-print":{"date-parts":[[2022,3,3]]},"abstract":"<jats:title>Abstract<\/jats:title>\n               <jats:p>Exposure of cultured mammalian cells to paraformaldehyde (PFA) is an effective approach to induce membrane blebs, which is followed by their detachment from the cellular cortex to yield giant membrane vesicles in extracellular spaces. Although PFA-induced giant vesicles have attracted significant interest in the field of cell membrane dynamics, their biochemical components and cytocompatibility remain largely unknown. In this report, we exposed human cervical cancer HeLa cells to PFA under metal-free buffer conditions to produce giant vesicles. We analyzed the components and structure of the purified PFA-induced giant vesicles. Co-culturing PFA-induced giant vesicles with exponentially growing HeLa cells resulted in docking of a significant number of the giant vesicles to the cell surface with seemingly no cytotoxicity. Intriguingly, we found that pre-treatment of HeLa cells with peptide-N-glycosidase and neuraminidase was effective in facilitating cellular uptake of constituents residing inside the vesicles. The results revealed further details about the effect of PFA on cell membranes and provide insights for studying the interaction between PFA-induced giant vesicles and cultured cells.<\/jats:p>","DOI":"10.1093\/jb\/mvab144","type":"journal-article","created":{"date-parts":[[2021,12,16]],"date-time":"2021-12-16T12:08:21Z","timestamp":1639656501000},"page":"339-347","source":"Crossref","is-referenced-by-count":3,"title":["Basic structure and cytocompatibility of giant membrane vesicles derived from paraformaldehyde-exposed human cells"],"prefix":"10.1093","volume":"171","author":[{"given":"Saya","family":"Okada","sequence":"first","affiliation":[{"name":"Department of Biological Science, Graduate School of Science and Technology, Kumamoto University, Kumamoto 860-8555, Japan"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Yuta","family":"Fukai","sequence":"additional","affiliation":[{"name":"Department of Biological Science, Graduate School of Science and Technology, Kumamoto University, Kumamoto 860-8555, Japan"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Yuki","family":"Tanoue","sequence":"additional","affiliation":[{"name":"International Research Center for Medical Sciences, Kumamoto University, Kumamoto 860-0811, Japan"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Hesham","family":"Nasser","sequence":"additional","affiliation":[{"name":"Divison of Molecular Virology and Genetics, Joint Research Center for Human Retrovirus Infection, Kumamoto University, Kumamoto 860-0811, Japan"},{"name":"Department of Clinical Pathology, Faculty of Medicine, Suez Canal University, Ismailia 41511, Egypt"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Takaichi","family":"Fukuda","sequence":"additional","affiliation":[{"name":"Department of Anatomy and Neurobiology, Graduate School of Medical Sciences, Kumamoto University, Kumamoto 860-8556, Japan"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Terumasa","family":"Ikeda","sequence":"additional","affiliation":[{"name":"Divison of Molecular Virology and Genetics, Joint Research Center for Human Retrovirus Infection, Kumamoto University, Kumamoto 860-0811, Japan"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Hisato","family":"Saitoh","sequence":"additional","affiliation":[{"name":"Department of Biological Science, Graduate School of Science and Technology, Kumamoto University, Kumamoto 860-8555, Japan"},{"name":"Faculty of Advanced Science and Technology (FAST), Kumamoto University, Kumamoto 860-8555, Japan"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"286","published-online":{"date-parts":[[2021,12,17]]},"reference":[{"key":"2022030711290061300_ref1","doi-asserted-by":"crossref","first-page":"8","DOI":"10.1017\/S1551929500057060","article-title":"Formaldehyde, formalin, paraformaldehyde and glutaraldehyde: what they are and what they do","volume":"8","author":"Kiernan","year":"2000","journal-title":"Micros. Today"},{"key":"2022030711290061300_ref2","doi-asserted-by":"crossref","DOI":"10.1101\/pdb.prot082594","article-title":"Formaldehyde cross-linking","volume":"2018","author":"Kim","year":"2018","journal-title":"Cold Spring Harb. Protoc."},{"key":"2022030711290061300_ref3","doi-asserted-by":"crossref","first-page":"299","DOI":"10.1007\/978-1-4939-8935-5_26","article-title":"An introduction to performing immunofluorescence staining","volume":"1897","author":"Im","year":"2019","journal-title":"Methods Mol. Biol."},{"key":"2022030711290061300_ref4","doi-asserted-by":"crossref","first-page":"3778","DOI":"10.1038\/s41467-019-11378-4","article-title":"The eleanor ncRNAs activate the topological domain of the ESR1 locus to balance against apoptosis","volume":"10","author":"Abdalla","year":"2019","journal-title":"Nat. Commun."},{"key":"2022030711290061300_ref5","doi-asserted-by":"crossref","first-page":"743","DOI":"10.1126\/science.982044","article-title":"Plasma membrane vesiculation: a new technique for isolation of plasma membranes","volume":"194","author":"Scott","year":"1976","journal-title":"Science"},{"key":"2022030711290061300_ref6","doi-asserted-by":"crossref","first-page":"229","DOI":"10.1242\/jcs.35.1.229","article-title":"Plasma membrane vesiculation in 3T3 and SV3T3 cells. I. Morphological and biochemical characterization","volume":"35","author":"Scott","year":"1979","journal-title":"J. Cell Sci."},{"key":"2022030711290061300_ref7","doi-asserted-by":"crossref","first-page":"245","DOI":"10.1242\/jcs.35.1.245","article-title":"Plasma membrane vesiculation in 3T3 and SV3T3 cells. II. Factors affecting the process of vesiculation","volume":"35","author":"Scott","year":"1979","journal-title":"J. Cell Sci."},{"key":"2022030711290061300_ref8","doi-asserted-by":"crossref","first-page":"1042","DOI":"10.1038\/nprot.2012.059","article-title":"Elucidating membrane structure and protein behavior using giant plasma membrane vesicles","volume":"7","author":"Sezgin","year":"2012","journal-title":"Nat. Protoc."},{"key":"2022030711290061300_ref9","doi-asserted-by":"crossref","first-page":"269","DOI":"10.1146\/annurev.biophys.32.110601.141803","article-title":"Model systems, lipid rafts, and cell membranes","volume":"33","author":"Simons","year":"2004","journal-title":"Annu. Rev. Biophys. Biomol. Struct."},{"key":"2022030711290061300_ref10","doi-asserted-by":"crossref","first-page":"3165","DOI":"10.1073\/pnas.0611357104","article-title":"Large-scale fluid\/fluid phase separation of proteins and lipids in giant plasma membrane vesicles","volume":"104","author":"Baumgart","year":"2007","journal-title":"Proc. Natl. Acad. Sci. USA."},{"key":"2022030711290061300_ref11","doi-asserted-by":"crossref","first-page":"25","DOI":"10.1016\/bs.ctm.2015.03.009","article-title":"Giant plasma membrane vesicles: models for understanding membrane organization","volume":"75","author":"Levental","year":"2015","journal-title":"Curr. Top. Membr."},{"key":"2022030711290061300_ref12","doi-asserted-by":"crossref","first-page":"194","DOI":"10.1016\/j.yexcr.2018.03.001","article-title":"The formation of giant plasma membrane vesicles enable new insights into the regulation of cholesterol efflux","volume":"365","author":"Sedgwick","year":"2018","journal-title":"Exp. Cell Res."},{"key":"2022030711290061300_ref13","doi-asserted-by":"crossref","first-page":"151","DOI":"10.1007\/978-1-4939-6358-4_11","article-title":"In situ SUMOylation and DeSUMOylation assays: fluorescent methods to visualize SUMOylation and DeSUMOylation in permeabilized cells","volume":"1475","author":"Yuasa","year":"2016","journal-title":"Methods Mol. Biol."},{"key":"2022030711290061300_ref14","doi-asserted-by":"crossref","first-page":"1124","DOI":"10.1016\/j.chom.2021.06.006","article-title":"SARS-CoV-2 spike L452R variant evades cellular immunity and increases infectivity","volume":"29","author":"Motozono","year":"2021","journal-title":"Cell Host Microbe."},{"key":"2022030711290061300_ref15","first-page":"14","article-title":"Three types of nuclear envelope assemblies associated with micronuclei","volume":"9","author":"Miyazaki","year":"2020","journal-title":"Cell Bio"},{"key":"2022030711290061300_ref16","doi-asserted-by":"crossref","first-page":"59","DOI":"10.1016\/j.ab.2018.07.012","article-title":"Wash-free instant detection of giant plasma membrane vesicles","volume":"557","author":"Okada","year":"2018","journal-title":"Anal. Biochem."},{"key":"2022030711290061300_ref17","doi-asserted-by":"crossref","first-page":"1137","DOI":"10.1007\/s10529-020-02848-7","article-title":"Chemical manipulations to facilitate membrane blebbing and vesicle shedding on the cellular cortex","volume":"42","author":"Okada","year":"2020","journal-title":"Biotechnol. Lett."},{"key":"2022030711290061300_ref18","doi-asserted-by":"crossref","first-page":"587","DOI":"10.1016\/j.cels.2017.05.009","article-title":"An optimized shotgun strategy for the rapid generation of comprehensive human proteomes","volume":"4","author":"Bekker-Jensen","year":"2017","journal-title":"Cell Syst."},{"key":"2022030711290061300_ref19","doi-asserted-by":"crossref","first-page":"428","DOI":"10.1016\/j.cell.2019.02.029","article-title":"Reassessment of exosome composition","volume":"177","author":"Jeppesen","year":"2019","journal-title":"Cell"},{"key":"2022030711290061300_ref20","doi-asserted-by":"crossref","first-page":"6401","DOI":"10.1021\/acs.biochem.7b00665","article-title":"Exosomes secreted by HeLa cells shuttle on their surface the plasma membrane-associated aialidase NEU3","volume":"56","author":"Paolini","year":"2017","journal-title":"Biochemistry"},{"key":"2022030711290061300_ref21","first-page":"6636","article-title":"Dendritic cells loaded with HeLa-derived exosomes simulate an antitumor immune response","volume":"15","author":"Ren","year":"2018","journal-title":"Oncol. Lett."},{"key":"2022030711290061300_ref22","doi-asserted-by":"crossref","first-page":"1722385","DOI":"10.1080\/20013078.2020.1722385","article-title":"Exosomes derived from HeLa cells break down vascular integrity by triggering endoplasmic reticulum stress in endothelial cells","volume":"9","author":"Lin","year":"2020","journal-title":"J. Extracell. Vesicles"},{"key":"2022030711290061300_ref23","first-page":"147","article-title":"Sugar-lectin interactions: how does wheat-germ agglutinin bind sialoglycoconjugates?","volume":"104","author":"Monsigny","year":"1980","journal-title":"FEBS J."},{"key":"2022030711290061300_ref24","doi-asserted-by":"crossref","first-page":"33","DOI":"10.1016\/B978-0-12-449945-4.50007-5","volume-title":"The Lectins: Properties, Functions and Applications in Biology and Medicine","author":"Goldstein","year":"1986"},{"key":"2022030711290061300_ref25","doi-asserted-by":"crossref","first-page":"1682","DOI":"10.1002\/cmdc.202000368","article-title":"ACE2, the receptor that enables infection by SARS-CoV-2: biochemistry, structure, allostery and evaluation of the potential development of ACE2 modulators","volume":"15","author":"Gross","year":"2020","journal-title":"Chem. Med. Chem."},{"key":"2022030711290061300_ref26","first-page":"2","article-title":"Evaluation of different PNGase F enzymes in immunoglobulin G and total plasma N-glycans analysis","volume":"31","author":"Vilaj","year":"2020","journal-title":"Glycobiology"},{"key":"2022030711290061300_ref27","doi-asserted-by":"crossref","first-page":"350","DOI":"10.1080\/10408347.2018.1531692","article-title":"Methods and current trends in determination of neuraminidase activity and evaluation of neuraminidase inhibitors","volume":"49","author":"H\u013easov\u00e1","year":"2019","journal-title":"Crit. Rev. Anal. Chem."},{"key":"2022030711290061300_ref28","doi-asserted-by":"crossref","first-page":"1757","DOI":"10.1016\/j.cell.2019.04.017","article-title":"Physical principles of membrane shape regulation by the glycocalyx","volume":"177","author":"Shurer","year":"2019","journal-title":"Cell"},{"key":"2022030711290061300_ref29","doi-asserted-by":"crossref","first-page":"346","DOI":"10.1038\/s41581-019-0129-4","article-title":"Glycosylation in health and disease","volume":"15","author":"Reily","year":"2019","journal-title":"Nat. Rev. Nephrol."},{"key":"2022030711290061300_ref30","doi-asserted-by":"crossref","DOI":"10.1016\/j.bbadis.2019.165608","article-title":"Mechanisms of extracellular vesicle uptake in stressed retinal pigment epithelial cell monolayers","volume":"1866","author":"Nicholson","year":"2020","journal-title":"Biochim. Biophys. Acta. Mol. Basis Dis."}],"container-title":["The Journal of Biochemistry"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/academic.oup.com\/jb\/advance-article-pdf\/doi\/10.1093\/jb\/mvab144\/42388681\/mvab144.pdf","content-type":"application\/pdf","content-version":"am","intended-application":"syndication"},{"URL":"https:\/\/academic.oup.com\/jb\/article-pdf\/171\/3\/339\/42748670\/mvab144.pdf","content-type":"application\/pdf","content-version":"vor","intended-application":"syndication"},{"URL":"https:\/\/academic.oup.com\/jb\/article-pdf\/171\/3\/339\/42748670\/mvab144.pdf","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2022,3,7]],"date-time":"2022-03-07T11:44:04Z","timestamp":1646653444000},"score":1,"resource":{"primary":{"URL":"https:\/\/academic.oup.com\/jb\/article\/171\/3\/339\/6464658"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2021,12,17]]},"references-count":30,"journal-issue":{"issue":"3","published-online":{"date-parts":[[2021,12,17]]},"published-print":{"date-parts":[[2022,3,3]]}},"URL":"https:\/\/doi.org\/10.1093\/jb\/mvab144","relation":{},"ISSN":["0021-924X","1756-2651"],"issn-type":[{"value":"0021-924X","type":"print"},{"value":"1756-2651","type":"electronic"}],"subject":[],"published-other":{"date-parts":[[2022,3]]},"published":{"date-parts":[[2021,12,17]]}}}