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SPHEROID CULTIVATION: AND MOTILITY REDUCTION THROUGH ACE2-RBDINTERACTION OF MONOLAYER COUNTERPARTS
Malmö University, Faculty of Health and Society (HS).
2021 (English)Independent thesis Basic level (degree of Bachelor), 10 credits / 15 HE creditsStudent thesisAlternative title
SFÄROIDODLING : OCH MOTILITETS STIMULERING GENOM ACE2-RBD INTERAKTION PÅ CELLER I MONOLAGER. (Swedish)
Abstract [en]

Spheroids are three-dimensional (3D) cell structures of aggregated cells that more closely mimic the cell-cell interactions and microenvironment of tumors. In this study 0,1% v/v of nano fibrillar cellulose hydrogel solubilized in spheroid medium was tested against static suspension culture. The aim was to determine whether either of the mediums was suitable to make the cell lines MCF-7, MDAMB-231, HMEC-1 as well as co-cultures of MCF-7/HMEC-1 and MDA-MB-231to form growth as spheroids. This study used holographic digital microscopy as well as light microscopy with image processing to analyze the cells. This study also examined the possible effect that the receptor-binding domain (RBD) of SARSCov2 might have on cell motility by interacting with angiotensin-converting enzyme 2 (ACE2). This study also included cross-linking of the RBD-ACE2complex to strengthen the RBD-ACE2 interaction. To this end, a motility assay was conducted on monolayers of the cell lines. This study found that among the cell lines and co-cultures only MCF-7 and MCF-7/HMEC-1 formed spheroids. The other cell lines and co-culture formed loose or tight aggregates. This study also showed that there was a significant difference in motility between the cell lines and indicated upon the difference between cells treated these results implicate decreased invasiveness by tumors in cancer patients due to toRBD-ACE2 interactions. This merits further experiments to determine whether-ACE2 treatment in vitro can serve to decrease the motility of tumor cells in cultures. 

Abstract [sv]

Sfäroider är tredimensionella (3D) cellstrukturer av aggregerade celler som närmare efterliknar cell-cell-interaktioner och mikromiljön hos tumörer. I denna studie undersöktes 0,1% v/v nanofibrillär cellulosa hydrogel löst sfäroidmedium mot statisk suspensionskultur. Syftet var att avgöra om något av substraten var lämpligt för att förmå cellinjerna MCF-7, MDA-MB-231, HMEC-1 samt cokultur av MCF-7 / HMEC-1 och MDA-MB-231 att bilda sfäroider. Denna studie använde holografisk digital mikroskopi samt ljusmikroskop med bild behandling för att analysera cellerna. Denna studie undersökte också eventuell effekt av receptorbindande domän (RBD) hos SARS-Cov2 på cell motilitet genom dessinteraktion med angiotensin konverterande enzym 2 (ACE2). I denna studieinkluderades också tvärbindning av ACE2-RBD komplexet syftet att stärka interaktionen mellan proteinerna. För detta ändamål genomfördes en motilitetsanalys på monolager av cellinjerna. Denna studie visade att bland cellinjerna och co-kulturerna var det endast MCF-7och MCF-7 / HMEC-1 som kunde odlas som sfäroider. De andra cellinjerna och co-kultur bildade lösa eller täta aggregat. Denna studie visade också att det fanns en signifikant skillnad i rörlighet mellan cellinjerna. Dessa resultat meriterar ytterligare experiment med RBD för att ta reda på omRBD-ACE2-stimulering kan användas till att minska motilitet hos tumörceller i in vitro kulturer då detta skulle kunna implicera minskad invasivitet av tumörer hos cancerpatienter.

Place, publisher, year, edition, pages
2021.
Keywords [en]
spheroid
Keywords [sv]
Sfäroid
National Category
Cell and Molecular Biology
Identifiers
URN: urn:nbn:se:mau:diva-49419OAI: oai:DiVA.org:mau-49419DiVA, id: diva2:1629458
Educational program
HS Biomedicinsk laboratorievetenskap
Presentation
2021-06-09, Zoom Meeting, https://mau-se.zoom.us/j/64565868879, 17:00
Supervisors
Examiners
Available from: 2022-11-09 Created: 2022-01-17 Last updated: 2022-11-09Bibliographically approved

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