A Comparison of Adipose-Derived Stem Cell Characteristics Between Quantum Expansion System and Flask-Based Culture
Author
Sørensen, Morten Brøndum
Term
4. term
Publication year
2023
Pages
20
Abstract
Chronic wounds persist when healing stalls in the inflammatory phase and substantially affect quality of life. Adipose-derived stem cells (ASCs) are a promising cell-based therapy, but reliable large-scale manufacturing is needed. This study compares ASCs expanded in a Quantum bioreactor with conventional flask-based culture, assessing surface marker profiles before and after freezing, stemness, and the ability to influence fibroblast and endothelial cell migration and proliferation. Using approaches including multicolor flow cytometry, tri-lineage differentiation, CFU and proliferation assays, and functional assays with conditioned media, the cells’ properties were evaluated. No meaningful differences were found between culture methods in stemness or in promoting fibroblast and endothelial growth. Some surface markers, including CD146, CD271, CD247, CD105, CD34, and CD31, differed between methods before and after freezing, but the freeze–thaw procedure itself did not markedly change overall marker profiles. Cells grown in flasks were significantly more viable than bioreactor-grown cells both before and after freezing. Overall, ASCs from the Quantum bioreactor behaved functionally similar to conventionally cultured ASCs, while flask culture preserved higher post-thaw viability, informing the choice of attachment materials for future large-scale clinical production.
Kroniske sår opstår, når sårheling går i stå i den inflammatoriske fase, og giver store gener for patienter. Adipøst-deriverede stamceller (ASCs) er lovende som cellebaseret behandling, men kræver pålidelig storskala-produktion. Dette studie sammenligner ASCs udvidet i en Quantum-bioreaktor med konventionel flaskebaseret dyrkning med fokus på overflademarkørprofil før og efter nedfrysning, stamcellekarakter (stemness) samt evnen til at påvirke fibroblasters og endotelcellers migration og proliferation. Ved hjælp af bl.a. multicolor-flowcytometri, tri-lineage-differentiering, CFU- og proliferationsanalyser samt funktionelle assays på konditioneret medie blev cellernes egenskaber vurderet. Der blev ikke fundet væsentlige forskelle mellem dyrkningsmetoderne i stamcelleegenskaber eller i evnen til at fremme fibroblast- og endotelcellevækst. Enkelte overflademarkører, herunder CD146, CD271, CD247, CD105, CD34 og CD31, varierede mellem dyrkningsmetoderne før og efter nedfrysning, men selve nedfrysning/optøningsproceduren ændrede ikke markant den overordnede markørprofil. Celler dyrket i kolber var signifikant mere levedygtige end celler fra bioreaktoren både før og efter nedfrysning. Samlet set opfører ASCs fra Quantum-bioreaktoren sig funktionelt på linje med konventionelt dyrkede ASCs, mens flaskebaseret dyrkning bevarer højere levedygtighed efter optøning, hvilket informerer valget af vedhæftningsmateriale til fremtidig storskala-produktion til klinisk brug.
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