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A concept for miniaturized 3-D cell culture using an extracellular matrix gel
KTH, Skolan för elektro- och systemteknik (EES), Mikrosystemteknik.ORCID-id: 0000-0002-3996-9279
KTH, Skolan för teknikvetenskap (SCI), Tillämpad fysik, Cellens fysik.
KTH, Skolan för elektro- och systemteknik (EES), Mikrosystemteknik.
KTH, Skolan för elektro- och systemteknik (EES), Mikrosystemteknik.ORCID-id: 0000-0001-9552-4234
Vise andre og tillknytning
2005 (engelsk)Inngår i: Electrophoresis, ISSN 0173-0835, E-ISSN 1522-2683, Vol. 26, nr 24, s. 4751-4758Artikkel i tidsskrift (Fagfellevurdert) Published
Abstract [en]

This paper presents a novel method to embed, anchor, and cultivate cells in a controlled 3-D flow-through microenvironment. This is realized using an etched silicon pillar flow chamber filled with extracellular matrix (ECM) gel mixed with cells. At 4 degrees C, while in liquid form, ECM gel is mixed with cells and injected into the chamber. Raising the temperature to 37 degrees C results in a gel, with cells embedded. The silicon pillars both stabilize and increase the surface to volume ratio of the gel. During polymerization the gel shrinks, thus creating channels, which enables perfusion through the chip. The pillars increase the mechanical stability of the gel permitting high surface flow rates without surface modifications. Within the structure cells were still viable and proliferating after 6 days of cultivation. Our method thus makes it possible to perform medium- to long-term cultivation of cells in a controlled 3-D environment. This concept opens possibilities to perform studies of cells in a more physiological environment compared to traditional 2-D cultures on flat substrates.

sted, utgiver, år, opplag, sider
2005. Vol. 26, nr 24, s. 4751-4758
Emneord [en]
embedded cells; extracellular matrix; microfluidics; three-dimensional cell culture
HSV kategori
Identifikatorer
URN: urn:nbn:se:kth:diva-11814DOI: 10.1002/elps.200500478ISI: 000234466300020Scopus ID: 2-s2.0-29644447617OAI: oai:DiVA.org:kth-11814DiVA, id: diva2:283449
Merknad
QC 20100723Tilgjengelig fra: 2009-12-28 Laget: 2009-12-28 Sist oppdatert: 2017-12-12bibliografisk kontrollert
Inngår i avhandling
1. Studies of Cellular Responses to External Stimuli in Engineered Microenvironments
Åpne denne publikasjonen i ny fane eller vindu >>Studies of Cellular Responses to External Stimuli in Engineered Microenvironments
2009 (engelsk)Doktoravhandling, med artikler (Annet vitenskapelig)
sted, utgiver, år, opplag, sider
Stockholm: KTH, 2009. s. xvi, 59
Serie
Trita-FYS, ISSN 0280-316X ; 09:73
HSV kategori
Identifikatorer
urn:nbn:se:kth:diva-11819 (URN)978-91-7415-529-7 (ISBN)
Disputas
2010-01-15, Sal FD5, AlbaNova, Roslagstullsbacken 21, Stockholm, 13:00 (engelsk)
Opponent
Veileder
Merknad
QC 20100806Tilgjengelig fra: 2009-12-29 Laget: 2009-12-29 Sist oppdatert: 2010-08-06bibliografisk kontrollert
2. MEMS Interfaces for Bioanalysis Systems
Åpne denne publikasjonen i ny fane eller vindu >>MEMS Interfaces for Bioanalysis Systems
2008 (engelsk)Doktoravhandling, med artikler (Annet vitenskapelig)
Abstract [en]

This thesis deals with various aspects of using open microfluidic interfaces. Three specific areas of application are studied.

The first is air-to-liquid interfacing in biosensors with possibilities for component inte­gration. A micromachined interface for airborne sample-to-liquid and droplet-to-liquid adsorption is discussed. It enables a robust sheet liquid flow serving as adsorption site. The inter­face properties are presented. Along with the interface, a novel method and system for rapid detection of dust and vapour-based narcotics and explosives traces is introduced. The QCM sensor detection principle with antibody immunoassay is described. Having shown the working principles of molecular adsorption to liquid surface and molecular detection with QCM technology, an integrated device is introduced. Diffusion as an effective transport mechanism in this microfluidic device is discussed. By holding the two components (inter­face and QCM) together with a double-sided adhesive, anisotropically vertically conductive tape, we achieve three functions, namely fixation, electrical connection and liquid sealing. Finally, enhanced electrostatic trapping of small particles to the liquid interface is demon­strated.

The second area concerns open microfluidics for the integration of capillary electropho­resis and mass spectroscopy. A technique for hyphenation between CE and MALDI-MS is presented. Two closed fused-silica capillaries were connected via a silicon chip comprising an open microcanal. The influence of the capillary-to-microcanal connection is discussed, as well as a simple technique to control evaporation from the open microcanal.

The third area concerns microfluidics enabling studies of single cells in asymmetric en­vironments. Using extracellular matrix or synthetic gel-embedding cells in an assay chamber, cells thrive and proliferate. This makes it possible to carry out medium to long term cultiva­tion of cells in a more physiological, controlled 3D environment than in traditional 2D cul­tures. The gels are discussed in terms of handling as well as their properties. A gel and micro­fluidic device for three dimensional cell culture with microgradient environments is pre­sented. Finally, a method for studying cilia-forming cells in asymmetric microfluidic environments is presented. Bending the primary cilium with a fluid flow will give rise to a response, but sensitivity to flow direction has only been sparsely studied. Design considerations are presented and discussed.

Abstract [sv]

Den här avhandlingen diskuterar olika aspekter av den öppna gränsytan hos styckevis öppna mikrofluidiksystem. Tre specifika användningsområden har studerats.

Det första är gränsytan mellan luft och vätska i en biosensor och de användningsområ­den som finns här. Ett mikrofabricerat interface för adsorption av luftburna substanser samt dropp-absorption diskuteras. Här används en rörlig vätskeyta som adsorbtionsyta och dess egenskaper presenteras. En ny metod för sprängämnes- och narkotikadetektering med interfa­cet introduceras. QCM-tekniken i kombination med antikroppskemi beskrivs. En integrerad lösning med dessa tekniker introduceras där diffusion utgör en effektiv transportmekanism. Med en dubbelsidig ledande tejp hålls komponenterna ihop, tätas och förses med ström. Slut­ligen presenteras elektrostatisk infångning av partiklar där den ena elektroden utgörs av väts­keytan.

Det andra området berör ett öppet mikrofluidiksystem för integrering av kapillärelek­trofores och masspektrometri. Teknik för att koppla ihop CE och MALDI-MS presente­ras. Två glaskapillärer har kopplats ihop med ett kiselchip med en öppen mikrokanal. Kopp­lingen mellan kapillären och chippet diskuteras liksom en enkel teknik för att kontrol­lera avdunstningen från den öppna mikrokanalen.

Det tredje området diskuterar hur mikrofluidik möjliggör studier av cellulära reaktioner i asymmetriska miljöer. Med inbäddning av celler i extracellulär matris eller syntetisk gel fås fysiologiskt relevant lokal miljö för celltillväxt och celldelning. Detta möjliggör studier av cellutveckling och cellreaktioner under lång tid i faktisk 3D-miljö till skillnad från den nuva­rande etablerade 2D-miljön. Gelerna diskuteras ur en hanteringssynpunkt liksom utifrån sina egenskaper. Ett system för cellodling i 3D med gradi­entmiljö presenteras och diskuteras. Slutligen presenteras ett system för studier av ciliefor­mande cellers respons där asymmetriska flöden ger upphov till böjning av cilier. Olika de­signaspekter diskuteras.

sted, utgiver, år, opplag, sider
Stockholm: KTH, 2008. s. xii, 56
Serie
Trita-EE, ISSN 1653-5146 ; 2008:002
Emneord
microfluidic interfacing, microfluidics, µTAS, sample transfer, biosensor, electronic nose, surface tension, quartz crystal microbalance, QCM, narcotics detection
HSV kategori
Identifikatorer
urn:nbn:se:kth:diva-4609 (URN)978-91-7178-846-7 (ISBN)
Disputas
2008-02-08, Sal M3, KTH, Brinellvägen 64, Stockholm, 10:00
Opponent
Veileder
Merknad
QC 20100927Tilgjengelig fra: 2008-01-18 Laget: 2008-01-18 Sist oppdatert: 2010-09-27bibliografisk kontrollert
3. Asymmetric Cellular Microenvironments
Åpne denne publikasjonen i ny fane eller vindu >>Asymmetric Cellular Microenvironments
2008 (engelsk)Licentiatavhandling, med artikler (Annet vitenskapelig)
Abstract [en]

This thesis presents methods to combine 3D cell culture, microfluidics and gradients on a controlled cellular scale. 3D cultures in biological extracellular matrix gels or synthetic gels bridge the gap between organ-tissue cultures and traditional 2D cultures. A device for embedding, anchoring and culturing cells in a controlled 3D flow through micro-environment was designed and evaluated. The device was realized using an etched silicon pillar flow chamber filled with gel mixed with cells. The pillars anchor and stabilize the gel as well as increase the surface to volume ratio, permitting higher surface flow rates and improving diffusion properties. Within the structure cells were still viable and proliferating after six days of cultivation, showing that it is possible to perform medium- to-long term cultivation of cells in a controlled 3D environment.

This concept was further developed to include controllable and time stable 3D microgradient environments. In this system stable diffusion gradients can be generated by the application of two parallel fluid flows with different composition against opposite sides of a gel plug with embedded cells. Culture for up to two weeks was performed showing cells still viable and proliferating. The cell tracer dye calcein was used to verify gradient formation as the fluorescent intensity in exposed cells was proportional to the position in the chamber. Cellular response to an applied stimulus was demonstrated by use of an adenosine triphosphate gradient where the onset of an intracellular calcium release also depends on cell position.

sted, utgiver, år, opplag, sider
Stockholm: KTH, 2008. s. vi, 33
Serie
Trita-FYS, ISSN 0280-316X ; 2007:87
HSV kategori
Identifikatorer
urn:nbn:se:kth:diva-4783 (URN)978-91-7178-848-1 (ISBN)
Presentation
(engelsk)
Veileder
Merknad
QC 20101119Tilgjengelig fra: 2008-06-02 Laget: 2008-06-02 Sist oppdatert: 2010-11-19bibliografisk kontrollert

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