Enhanced bile canaliculi formation enabling direct recovery of biliary metabolites of hepatocytes in 3D collagen gel microcavities

Lab on a Chip ◽  
2012 ◽  
Vol 12 (10) ◽  
pp. 1857 ◽  
Author(s):  
Hitoshi Matsui ◽  
Shoji Takeuchi ◽  
Tomoharu Osada ◽  
Teruo Fujii ◽  
Yasuyuki Sakai
Keyword(s):  
Collagen Gel ◽  
Bile Canaliculi ◽  
3D Collagen ◽  
Biliary Metabolites

scholarly journals Extending In-Plane Impedance Measurements from 2D to 3D Cultures: Design Considerations

2021 ◽  
Vol 8 (1) ◽  
pp. 11
Author(s):  
Sorel E. De Leon ◽  
Lana Cleuren ◽  
Zay Yar Oo ◽  
Paul R. Stoddart ◽  
Sally L. McArthur
Keyword(s):  
Three Dimensional ◽  
3D Culture ◽  
Collagen Gel ◽  
Impedance Measurement ◽  
Impedance Spectrum ◽  
3D Models ◽  
Low Frequencies ◽  
3D Cell Cultures ◽  
3D Collagen ◽  
2D And 3D

Three-dimensional (3D) cell cultures have recently emerged as tools for biologically modelling the human body. As 3D models make their way into laboratories there is a need to develop characterisation techniques that are sensitive enough to monitor the cells in real time and without the need for chemical labels. Impedance spectroscopy has been shown to address both of these challenges, but there has been little research into the full impedance spectrum and how the different components of the system affect the impedance signal. Here we investigate the impedance of human fibroblast cells in 2D and 3D collagen gel cultures across a broad range of frequencies (10 Hz to 5 MHz) using a commercial well with in-plane electrodes. At low frequencies in both 2D and 3D models it was observed that protein adsorption influences the magnitude of the impedance for the cell-free samples. This effect was eliminated once cells were introduced to the systems. Cell proliferation could be monitored in 2D at intermediate frequencies (30 kHz). However, the in-plane electrodes were unable to detect any changes in the impedance at any frequency when the cells were cultured in the 3D collagen gel. The results suggest that in designing impedance measurement devices, both the nature and distribution of the cells within the 3D culture as well as the architecture of the electrodes are key variables.

scholarly journals Compression Induced Stress Response of Nucleus Pulposus Cells in 3D Collagen Gel

2016 ◽  
Vol 6 (1_suppl) ◽  
pp. s-0036-1582596-s-0036-1582596
Author(s):  
Wai Hon Chooi ◽  
Barbara P. Chan
Keyword(s):  
Stress Response ◽  
Nucleus Pulposus ◽  
Collagen Gel ◽  
Nucleus Pulposus Cells ◽  
Induced Stress ◽  
3D Collagen

scholarly journals ROCK-generated contractility regulates breast epithelial cell differentiation in response to the physical properties of a three-dimensional collagen matrix

2003 ◽  
Vol 163 (3) ◽  
pp. 583-595 ◽  
Author(s):  
Michele A. Wozniak ◽  
Radhika Desai ◽  
Patricia A. Solski ◽  
Channing J. Der ◽  
Patricia J. Keely
Keyword(s):  
Epithelial Cells ◽  
Three Dimensional ◽  
Collagen Gel ◽  
Collagen Matrix ◽  
Breast Epithelial Cell ◽  
Collagen Gels ◽  
Culture Dish ◽  
Breast Epithelial Cells ◽  
3D Collagen ◽  
Breast Epithelial

Breast epithelial cells differentiate into tubules when cultured in floating three-dimensional (3D) collagen gels, but not when the cells are cultured in the same collagen matrix that is attached to the culture dish. These observations suggest that the biophysical properties of collagenous matrices regulate epithelial differentiation, but the mechanism by which this occurs is unknown. Tubulogenesis required the contraction of floating collagen gels through Rho and ROCK-mediated contractility. ROCK-mediated contractility diminished Rho activity in a floating 3D collagen gel, and corresponded to a loss of FAK phosphorylated at Y397 localized to 3D matrix adhesions. Increasing the density of floating 3D collagen gels also disrupted tubulogenesis, promoted FAK phosphorylation, and sustained high Rho activity. These data demonstrate the novel finding that breast epithelial cells sense the rigidity or density of their environment via ROCK-mediated contractility and a subsequent down-regulation of Rho and FAK function, which is necessary for breast epithelial tubulogenesis to occur.

scholarly journals Integrated computational and experimental pipeline for quantifying local cell–matrix interactions

2021 ◽  
Vol 11 (1) ◽  
Author(s):  
Hugh Xiao ◽  
Ryan Y. Nguyen ◽  
Ryan LaRanger ◽  
Erica L. Herzog ◽  
Michael Mak
Keyword(s):  
Collagen Gel ◽  
Collagen Matrix ◽  
Cellular Level ◽  
Lung Fibroblasts ◽  
Cellular Interactions ◽  
Cell Matrix ◽  
Matrix Interactions ◽  
3D Collagen ◽  
Local Cell ◽  
Cell Matrix Interactions

AbstractCellular interactions with the extracellular matrix (ECM) play a key role in modulating biological processes. While studies have identified key molecular factors of these interactions, the mechanical regulation associated with these interactions is not well characterized. To address this, we present an image analysis platform to analyze time-dependent dynamics observed in lung fibroblasts embedded in a 3D collagen matrix. Combining drug studies with quantitative analysis of cell–matrix interactions, our results are able to provide cellular level quantitative insights for mechanical and biophysical phenomena relevant to cell-ECM interactions. This system overall represents an initial pipeline for understanding cell mechanics in a 3D collagen gel and their implications in a physiologically relevant context.

Guided Axon Growth by Gradients of Adhesion in Collagen Gels

2008 ◽  
Author(s):  
Harini G. Sundararaghavan ◽  
Gary A. Monteiro ◽  
David I. Shreiber
Keyword(s):  
Spinal Cord ◽  
Collagen Gel ◽  
Axon Growth ◽  
Neurite Growth ◽  
Gradient System ◽  
Spinal Cord Regeneration ◽  
Collagen Gels ◽  
Direct Growth ◽  
3D Collagen

During development, neurites are directed by gradients of attractive and repulsive soluble (chemotactic) cues and substrate-bound adhesive (haptotactic) cues. Many of these cues have been extensively researched in vitro, and incorporated into strategies for nerve and spinal cord regeneration, primarily to improve the regenerative environment. To enhance and direct growth, we have developed a system to create 1D gradients of adhesion through a 3D collagen gel using microfluidics. We test our system using collagen grafted with bioactive peptide sequences, IKVAV and YIGSR, from laminin — an extra-cellular matrix (ECM) protein known to strongly influence neurite outgrowth. Gradients are established from ∼0.37mg peptide/mg collagen – 0, and ∼0.18 mg peptide/mg collagen – 0 of each peptide and tested using chick dorsal root ganglia (DRG). Neurite growth is evaluated 5 days after gradient formation. Neurites show increased growth in the gradient system when compared to control and biased growth up the gradient of peptides. Growth in YIGSR-grafted collagen increased with steeper gradients, whereas growth in IKVAV-grafted collagen decreased with steeper gradients. These results demonstrate that neurite growth can be enhanced and directed by controlled, immobilized, haptotactic gradients through 3D scaffolds, and suggest that including these gradients in regenerative therapies may accelerate nerve and spinal cord regeneration.

Microfluidic Generation of Adhesion Gradients Through 3D Collagen Gels: Implications for Neural Tissue Engineering

2008 ◽  
Author(s):  
Harini G. Sundararaghavan ◽  
Gary A. Monteiro ◽  
David I. Shreiber
Keyword(s):  
Spinal Cord ◽  
Collagen Gel ◽  
Neurite Growth ◽  
Gradient System ◽  
Spinal Cord Regeneration ◽  
Collagen Gels ◽  
Direct Growth ◽  
3D Collagen ◽  
Regenerative Therapies

During development, neurites are directed by gradients of attractive and repulsive soluble (chemotactic) cues and substrate-bound adhesive (haptotactic) cues. Many of these cues have been extensively researched in vitro, and incorporated into strategies for nerve and spinal cord regeneration, primarily to improve the regenerative environment. To enhance and direct growth, we have developed a system to create 1D gradients of adhesion through a 3D collagen gel using microfluidics. We test our system using collagen grafted with bioactive peptide sequences, IKVAV and YIGSR, from laminin — an extra-cellular matrix (ECM) protein known to strongly influence neurite outgrowth [1, 2]. Gradients are established from 0.14 mg/ml–0, and 0.07 mg/ml–0 of each peptide and tested using chick dorsal root ganglia (DRG). Neurite growth is evaluated 5 days after gradient formation. Neurites show increased growth in the gradient system when compared to control and biased growth up the gradient of peptides. These results demonstrate that neurite growth can be enhanced and directed by controlled, immobilized, haptotactic gradients through 3D scaffolds, and suggest that including these gradients in regenerative therapies may accelerate nerve and spinal cord regeneration.

scholarly journals Cellular Fragments as Biomaterial for Rapid In Vitro Bone-Like Tissue Synthesis

2020 ◽  
Vol 21 (15) ◽  
pp. 5327
Author(s):  
Mst Nahid Akhter ◽  
Emilio Satoshi Hara ◽  
Koichi Kadoya ◽  
Masahiro Okada ◽  
Takuya Matsumoto
Keyword(s):  
Collagen Gel ◽  
Culture Conditions ◽  
Substrate Material ◽  
Dependent Manner ◽  
Concentration Dependent Manner ◽  
Freeze Dried ◽  
Ionic Detergent ◽  
3D Collagen ◽  
Fetal Bovine

Current stem cell-based techniques for bone-like tissue synthesis require at least two to three weeks. Therefore, novel techniques to promote rapid 3D bone-like tissue synthesis in vitro are still required. In this study, we explored the concept of using cell nanofragments as a substrate material to promote rapid bone formation in vitro. The methods for cell nanofragment fabrication were ultrasonication (30 s and 3 min), non-ionic detergent (triton 0.1% and 1%), or freeze-dried powder. The results showed that ultrasonication for 3 min allowed the fabrication of homogeneous nanofragments of less than 150 nm in length, which mineralized surprisingly in just one day, faster than the fragments obtained from all other methods. Further optimization of culture conditions indicated that a concentration of 10 mM or 100 mM of β-glycerophosphate enhanced, whereas fetal bovine serum (FBS) inhibited in a concentration-dependent manner, the mineralization of the cell nanofragments. Finally, a 3D collagen-cell nanofragment-mineral complex mimicking a bone-like structure was generated in just two days by combining the cell nanofragments in collagen gel. In conclusion, sonication for three min could be applied as a novel method to fabricate cell nanofragments of less than 150 nm in length, which can be used as a material for in vitro bone tissue engineering.

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