Time Lapse Monitoring of CaCo-2 Cell Shapes and Shape Dependence of the Distribution of Integrin β1 and F-Actin on their Basal Membrane

Jokhadar, Špela Zemljič; Šuštar, Vid; Svetina, S.; Batista, Urška

doi:10.1080/15419060902957296

Cited by 9 publications

(5 citation statements)

References 27 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…Caco-2 cells were observed to adhere onto the ECM coated Teflon membrane within 30 min after cell seeding. During the phase of active growth, it was observed that once the flow of cell culture media was started (first 24 hrs of cell culture), the Caco-2 cells spread out [ 57 ], although, in some regions the cells grew in clusters ( Fig 4A ). It was also observed that flowing DMEM at 0.5μl/ml for the first 16-24hrs across the cells, was sufficient to wash-out the unattached Caco-2 cells ( S6B Fig ) without releasing the cells.…”

Section: Resultsmentioning

confidence: 99%

A multi-chamber microfluidic intestinal barrier model using Caco-2 cells for drug transport studies

et al. 2018

View full text Add to dashboard Cite

This paper presents the design and fabrication of a multi-layer and multi-chamber microchip system using thiol-ene ‘click chemistry’ aimed for drug transport studies across tissue barrier models. The fabrication process enables rapid prototyping of multi-layer microfluidic chips using different thiol-ene polymer mixtures, where porous Teflon membranes for cell monolayer growth were incorporated by masked sandwiching thiol-ene-based fluid layers. Electrodes for trans-epithelial electrical resistance (TEER) measurements were incorporated using low-melting soldering wires in combination with platinum wires, enabling parallel real-time monitoring of barrier integrity for the eight chambers. Additionally, the translucent porous Teflon membrane enabled optical monitoring of cell monolayers. The device was developed and tested with the Caco-2 intestinal model, and compared to the conventional Transwell system. Cell monolayer differentiation was assessed via in situ immunocytochemistry of tight junction and mucus proteins, P-glycoprotein 1 (P-gp) mediated efflux of Rhodamine 123, and brush border aminopeptidase activity. Monolayer tightness and relevance for drug delivery research was evaluated through permeability studies of mannitol, dextran and insulin, alone or in combination with the absorption enhancer tetradecylmaltoside (TDM). The thiol-ene-based microchip material and electrodes were highly compatible with cell growth. In fact, Caco-2 cells cultured in the device displayed differentiation, mucus production, directional transport and aminopeptidase activity within 9–10 days of cell culture, indicating robust barrier formation at a faster rate than in conventional Transwell models. The cell monolayer displayed high TEER and tightness towards hydrophilic compounds, whereas co-administration of an absorption enhancer elicited TEER-decrease and increased permeability similar to the Transwell cultures. The presented cell barrier microdevice constitutes a relevant tissue barrier model, enabling transport studies of drugs and chemicals under real-time optical and functional monitoring in eight parallel chambers, thereby increasing the throughput compared to previously reported microdevices.

show abstract

Section: Resultsmentioning

confidence: 99%

A multi-chamber microfluidic intestinal barrier model using Caco-2 cells for drug transport studies

et al. 2018

View full text Add to dashboard Cite

show abstract

“…B). Although other TLVM studies document live Caco2 cell phenomena , they often indicate merely that the cells were incubated on a heated stage or in a chamber at 37 degrees Celsius and do not demonstrate that the cells themselves experience a physiological environment and behave as expected under physiological conditions. Our device measures conditions in the immediate environment surrounding the cells and recapitulates the pH, humidity, and cell growth rate maintained by a standard incubator.…”

Section: Discussionmentioning

confidence: 99%

Novel aspects of live intestinal epithelial cell function revealed using a custom time‐lapse video microscopy apparatus

Papetti

Kozlowski

2018

Cytometry Pt A

View full text Add to dashboard Cite

Many aspects of cell physiology, including migration, membrane function, and cell division, are best understood by observing live cell dynamics over time using video microscopy. To probe these phenomena in colon epithelial cells using simple components with a limited budget, we have constructed an inexpensive (<$410) self-contained apparatus, consisting of a closed-loop, feedback-controlled system regulated by a PID (proportional-integrative-derivative) controller contained within a 0.077 m insulated acrylic box. Temperature, humidity, pH, and proliferative capacity of colon epithelial cells in this system mimic those in a standard tissue culture incubator for over four days. Our system offers significant advantages over existing cost-prohibitive commercially available and custom-made devices because of its very low cost, use of PID temperature control, lack of reliance on constant infusion of external humidified, heated air or carbon dioxide, ability to directly measure cell culture medium temperature, and combination of exquisite cellular detail with minimal focus drift under physiological conditions for extended periods of time. Using this apparatus, coupled with an inverted microscope equipped with phase contrast optics and a programmable digital camera, we have observed many events in colon epithelial cells not visible by static imaging, including kinetics of normal and abnormal mitoses, dynamic membrane structures, intracellular vesicle movements, and cell migration. © 2018 International Society for Advancement of Cytometry.

show abstract

“…The advantages of ECM membranes in cell culture were demonstrated by Wang et al, who compared colonic cell culture on three different fibronectin-coated devices. A membrane-less device showed a decline in cell viability after Day 1 and maintained round cell morphology, which indicates a lack of differentiation 139 . The viability further decreased to 76% on Day 5, indicating an early apoptotic phase 136 .…”

Section: Biological Ecm Protein Membranesmentioning

confidence: 98%

Polymeric and biological membranes for organ-on-a-chip devices

Corral-Nájera,

Chauhan,

Serna-Saldívar

et al. 2023

Microsyst Nanoeng

View full text Add to dashboard Cite

Membranes are fundamental elements within organ-on-a-chip (OOC) platforms, as they provide adherent cells with support, allow nutrients (and other relevant molecules) to permeate/exchange through membrane pores, and enable the delivery of mechanical or chemical stimuli. Through OOC platforms, physiological processes can be studied in vitro, whereas OOC membranes broaden knowledge of how mechanical and chemical cues affect cells and organs. OOCs with membranes are in vitro microfluidic models that are used to replace animal testing for various applications, such as drug discovery and disease modeling. In this review, the relevance of OOCs with membranes is discussed as well as their scaffold and actuation roles, properties (physical and material), and fabrication methods in different organ models. The purpose was to aid readers with membrane selection for the development of OOCs with specific applications in the fields of mechanistic, pathological, and drug testing studies. Mechanical stimulation from liquid flow and cyclic strain, as well as their effects on the cell’s increased physiological relevance (IPR), are described in the first section. The review also contains methods to fabricate synthetic and ECM (extracellular matrix) protein membranes, their characteristics (e.g., thickness and porosity, which can be adjusted depending on the application, as shown in the graphical abstract), and the biological materials used for their coatings. The discussion section joins and describes the roles of membranes for different research purposes and their advantages and challenges.

show abstract

Time Lapse Monitoring of CaCo-2 Cell Shapes and Shape Dependence of the Distribution of Integrin β1 and F-Actin on their Basal Membrane

Cited by 9 publications

References 27 publications

A multi-chamber microfluidic intestinal barrier model using Caco-2 cells for drug transport studies

A multi-chamber microfluidic intestinal barrier model using Caco-2 cells for drug transport studies

Novel aspects of live intestinal epithelial cell function revealed using a custom time‐lapse video microscopy apparatus

Polymeric and biological membranes for organ-on-a-chip devices

Contact Info

Product

Resources

About