Building a microfluidic cell culture platform with stiffness control using Loctite 3525 glue

Vázquez‐Victorio, Genaro; Peto-Gutiérrez, Cindy; Díaz-Bello, Beatriz; Cano-Jorge, Mariel; Pérez-Calixto, Daniel; Jiménez‐Escobar, Alejandra; Espinosa‐Matías, Silvia; Martínez, Reyna Lara; Courson, Rémi; Malaquin, Laurent; Zamarrón-Hernández, Diego; Hautefeuille, Mathieu

doi:10.1039/c9lc00649d

Cited by 10 publications

(6 citation statements)

References 56 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…It was recently reported that mouse primary hepatocytes respond to stiff PAA hydrogels (with an elastic modulus ≥10 kPa) by doubling their area in the first 12 h after seeding; however, this effect seems to be attenuated in viscoelastic PAA HGs [26,37]. To evaluate our conditions and confirm previous results, we decided to use soft (1 kPa, elastic modulus) and stiff (20 kPa, elastic modulus) polyacrylamide hydrogels conjugated with rat tail collagen type I as described in [74]. We evaluated the spreading of primary hepatocytes at 24 and 72 h after culture (Figure 1A).…”

Section: Stiffness Promotes Cell Aggregation and Triggers Spreading In Primary Hepatocytes Whereas Collagen Type I Promotes Cell Adhesionmentioning

confidence: 60%

“…Immunofluorescence and terminal deoxynucleotidyl transferase dUTP nick end labeling (TUNEL) assays were performed in rat primary hepatocytes cultured on PAA HGs as described in [74]. Briefly, hepatocytes were washed once with pre-warmed 1× DPBS and immediately fixed with 4% paraformaldehyde (PFA) in DPBS 1× at 37 • C for 20 min.…”

Section: Immunofluorescence and Tunel Assaymentioning

confidence: 99%

See 1 more Smart Citation

Fibrillar Collagen Type I Participates in the Survival and Aggregation of Primary Hepatocytes Cultured on Soft Hydrogels

et al. 2020

Self Cite

View full text Add to dashboard Cite

Liver is an essential organ that carries out multiple functions such as glycogen storage, the synthesis of plasma proteins, and the detoxification of xenobiotics. Hepatocytes are the parenchyma that sustain almost all the functions supported by this organ. Hepatocytes and non-parenchymal cells respond to the mechanical alterations that occur in the extracellular matrix (ECM) caused by organogenesis and regenerating processes. Rearrangements of the ECM modify the composition and mechanical properties that result in specific dedifferentiation programs inside the hepatic cells. Quiescent hepatocytes are embedded in the soft ECM, which contains an important concentration of fibrillar collagens in combination with a basement membrane-associated matrix (BM). This work aims to evaluate the role of fibrillar collagens and BM on actin cytoskeleton organization and the function of rat primary hepatocytes cultured on soft elastic polyacrylamide hydrogels (PAA HGs). We used rat tail collagen type I and Matrigel® as references of fibrillar collagens and BM respectively and mixed different percentages of collagen type I in combination with BM. We also used peptides obtained from decellularized liver matrices (dECM). Remarkably, hepatocytes showed a poor adhesion in the absence of collagen on soft PAA HGs. We demonstrated that collagen type I inhibited apoptosis and activated extracellular signal-regulated kinases 1/2 (ERK1/2) in primary hepatocytes cultured on soft hydrogels. Epidermal growth factor (EGF) was not able to rescue cell viability in conjugated BM but affected cell aggregation in soft PAA HGs conjugated with combinations of different proportions of collagen and BM. Interestingly, actin cytoskeleton was localized and preserved close to plasma membrane (cortical actin) and proximal to intercellular ducts (canaliculi-like structures) in soft conditions; however, albumin protein expression was not preserved, even though primary hepatocytes did not remodel their actin cytoskeleton significantly in soft conditions. This investigation highlights the important role of fibrillar collagens on soft hydrogels for the maintenance of survival and aggregation of the hepatocytes. Data suggest evaluating the conditions that allow the establishment of optimal biomimetic environments for physiology and cell biology studies, where the phenotype of primary cells may be preserved for longer periods of time.

show abstract

Section: Stiffness Promotes Cell Aggregation and Triggers Spreading In Primary Hepatocytes Whereas Collagen Type I Promotes Cell Adhesionmentioning

confidence: 60%

Section: Immunofluorescence and Tunel Assaymentioning

confidence: 99%

Fibrillar Collagen Type I Participates in the Survival and Aggregation of Primary Hepatocytes Cultured on Soft Hydrogels

et al. 2020

Self Cite

View full text Add to dashboard Cite

show abstract

“…When the determination of Young's modulus of a material is required, for instance for quantifying the influence of substrate mechanics on cell spreading [62,63] or determining forces reliably using traction force microscopy measurements [64], our method also readily enables its determination from the relaxation data and GMM analysis. For that, we compared the calculated values of the elastic modulus obtained with conventional microindentation and with the relaxation tests.…”

Section: Comparison Between Microindentation and Relaxation Mechanicamentioning

confidence: 99%

Determination by Relaxation Tests of the Mechanical Properties of Soft Polyacrylamide Gels Made for Mechanobiology Studies

et al. 2021

Self Cite

View full text Add to dashboard Cite

Following the general aim of recapitulating the native mechanical properties of tissues and organs in vitro, the field of materials science and engineering has benefited from recent progress in developing compliant substrates with physical and chemical properties similar to those of biological materials. In particular, in the field of mechanobiology, soft hydrogels can now reproduce the precise range of stiffnesses of healthy and pathological tissues to study the mechanisms behind cell responses to mechanics. However, it was shown that biological tissues are not only elastic but also relax at different timescales. Cells can, indeed, perceive this dissipation and actually need it because it is a critical signal integrated with other signals to define adhesion, spreading and even more complicated functions. The mechanical characterization of hydrogels used in mechanobiology is, however, commonly limited to the elastic stiffness (Young’s modulus) and this value is known to depend greatly on the measurement conditions that are rarely reported in great detail. Here, we report that a simple relaxation test performed under well-defined conditions can provide all the necessary information for characterizing soft materials mechanically, by fitting the dissipation behavior with a generalized Maxwell model (GMM). The simple method was validated using soft polyacrylamide hydrogels and proved to be very useful to readily unveil precise mechanical properties of gels that cells can sense and offer a set of characteristic values that can be compared with what is typically reported from microindentation tests.

show abstract

“…Part Ba adapted with permission from ref. 77 substrates in the OoC device, whereby the stiffness 190,191 or surface topographies 192,193 of the substrates can be precisely controlled.…”

Section: Probing and Mimicking The Cell Microenvironmentmentioning

confidence: 99%

A guide to the organ-on-a-chip

Leung

Haan

Ronaldson-Bouchard

et al. 2022

Nat Rev Methods Primers

421

276

View full text Add to dashboard Cite

The organ-on-a-chip (OoC) is an intriguing scientific and technological development in which biology is coupled with microtechnology 1,2 to mimic key aspects of human physiology. The chip takes the form of a microfluidic device containing networks of hair-fine microchannels for guiding and manipulating minute volumes (picolitres up to millilitres) of solution [3][4][5] . The organ is a more relatable term that refers to the miniature tissues grown and residing in the microfluidic chips, which can recapitulate one or more tissue-specific functions. Although they are much simpler than native tissues and organs, scientists have discovered that these systems can often serve as effective mimics of human physiology and disease. OoCs comprise advanced in vitro technology that enables experimentation with biological cells and tissues outside the body. This is achieved by containing them inside vessels conditioned to sustain a reasonable semblance of the in vivo environment, from a biochemical and physical point of view. Working on the microscale lends a unique opportunity to attain a higher level of control over the microenvironment that ensures tissue life support, as well as a means to directly observe cell and tissue behaviour.The OoC is a relatively recent addition to the toolbox of model biological systems available to life science researchers to probe aspects of human pathophysiology and disease. These systems cover a spectrum of physiological relevance, with 2D cell cultures the least relevant, followed in increasing order by 3D cell cultures, organoids and OoCs. Unsurprisingly, the use of model organisms such as mice and Drosophila physiologically exceeds engineered tissue approaches 6,7 . While biological complexity increases with physiological relevance in model organisms, this unfortunately leads to increased experimental difficulty. In vivo physiological processes are, in many ways, the least accessible to direct investigation in mice, humans and other mammals, despite significant advances in in vivo imaging. However, 2D and 3D cell cultures, such as spheroids and stem cell-derived organoids, sacrifice some aspects of in vivo relevance to facilitate experimentation. The OoC may be regarded as a bridging technology, offering the ability to work with complex cell cultures, while providing better engineered microenvironments to maximize the model.Following on from early concepts, including animal-on-a-chip 8 , body-on-a-chip 9 and breathing lung-on-a-chip 10 , research in the OoC and microphysiological systems fields has grown exponentially; evidenced by numerous excellent reviews published recently 1,2,11 . Recognition of OoC technology now extends far beyond university laboratories, driven by a need to better understand the human physiology underlying health and disease, and to find new approaches to improve the human condition. The World Economic Forum, for instance, selected the OoC as one of the top ten emerging technologies in 2016 (ref. 12

show abstract

Building a microfluidic cell culture platform with stiffness control using Loctite 3525 glue

Abstract: The study of cell response to mechanotransduction signals requires designing culture substrates offering biocompatibility and adhesion, stiffness control and dynamics, patternability at microscale and integration in microfluidics chips.

Cited by 10 publications

References 56 publications

Fibrillar Collagen Type I Participates in the Survival and Aggregation of Primary Hepatocytes Cultured on Soft Hydrogels

Fibrillar Collagen Type I Participates in the Survival and Aggregation of Primary Hepatocytes Cultured on Soft Hydrogels

Determination by Relaxation Tests of the Mechanical Properties of Soft Polyacrylamide Gels Made for Mechanobiology Studies

A guide to the organ-on-a-chip

Contact Info

Product

Resources

About