Hemmingsen, Mette

Mikrofluid celledyrkning har det sidste årti været et forskningsområde med stor opmærksomhed på grund af dets potentiale for nøjere at kunne efterligne de in vivo cellulære omgivelser i forhold til hvad, der er muligt ved konventionelle celledyrkningsmetoder. Mange spændende og komplekse devices, som giver mulighed for, for eksempel, præcis kontrol af de kemiske omgivelser, 3D kulturer, kontrolleret samdyrkning af forskellige typer celler eller automatiseret individuel kontrol af op til 96 celledyrkningskamre i ét integreret system, er blevet præsenteret. Til trods for de storartede nye muligheder for at udføre hidtil ukendte forsøgsopsætninger mangler disse devices stadig at blive generelt implementeret i biologiske forskningslaboratorier.I dette projekt er brugervenlighed og anvendelighed af mikrofluidiske celledyrkningssystemer blevet undersøgt. De testede systemer udviser gode egenskaber angående optik og kompatibilitet med standard laboratorieudstyr og procedurer. Sådanne meget brugervenlige systemer er opnået ved integration af fluid aktivering, væskereservoirer, væskeforbindelser og celledyrkningskamre i en enkelt transportabel enhed. Imidlertid er det vist at forbedringer angående robusthed af individuelle systemdele og dermed driftsikkerhed af systemerne, er nødvendig. Dette er muligvis en af grundene til den manglende implementering af mikrofluide celledyrkningssystemer i biologiske forskningslaboratorier.Procedurer for at udføre langtids-mikrofluide perfusions celledyrkningseksperimenter er blevet fastsat. Ydermere er en succesfuld anvendelse af mikrofluid perfusions celledyrkningssystemet vist ved undersøgelse af fedt-afledte stamcellers differentiering til fedtceller, hvor vi har afsløret at parakrin/autokrin signalering er involveret i differentiering af en fedtstamcellepopulation til fedtceller. Vi har dermed demonstreret at mikrofluid perfusions celledyrkningssystemer er et mere kraftfuldt og brugbart redskab til at undersøge parakrin/autokrin signalering indenfor en cellepopulation i forhold til konventionel statisk celledyrkning. Vi har således været i stand til at udføre ny biologisk forskning ved brug af vores udviklede mikrofluid perfusions celledyrkningssystemer.Microfluidic cell culture has been a research area with great attention the last decade due to its potential to mimic the in vivo cellular environment more closely compared to what is possible by conventional cell culture methods. Many exciting and complex devices have been presented providing possibilities for, for example, precise control of the chemical environment, 3D cultures, controlled co-culture of different cell types or automated, individual control of up to 96 cell culture chambers in one integrated system. Despite the great new opportunities to perform novel experimental designs, these devices still lack general implementation into biological research laboratories.In this project, the usability and applicability of microfluidic cell culture systems have been investigated. The tested systems display good properties regarding optics and compatibility with standard laboratory equipment and procedures. Such highly usable systems were achieved by integration of fluidic actuation, liquid reservoirs, liquid interconnections and cell culture chambers in a single portable unit. However, improvements regarding robustness of individual system parts and thereby reliability of the systems are shown to be needed. This is possibly one of the reasons for the lack of implementation of microfluidic cell culture systems into biological research laboratories.Procedures to perform long-term microfluidic perfusion cell culture experiments have been established. Furthermore, successful application of the microfluidic perfusion cell culture system is shown by investigation of adipose-derived stem cell (ASC) differentiation into adipocytes, where we have revealed that paracrine/autocrine signaling is involved in differentiation of a population of ASCs into adipocytes. We have thereby demonstrated that microfluidic perfusion cell culture systems are a more powerful and a useful tool to investigate paracrine/autocrine signaling within a cell population compared to conventional static cell culture. Thus, we have been able to perform novel biological research by the use of our developed microfluidic perfusion cell culture systems

Online Research Database In Technology

Usability and Applicability of Microfluidic Cell Culture Systems

http://orbit.dtu.dk/ws/files/51216779/Thesis_Final_version..PDF

Usability and Applicability of Microfluidic Cell Culture Systems

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Online Research Database In Technology