scholarly journals Characterizing Cell Migration Within Three-dimensional In Vitro Wound Environments

10.3791/56099 ◽  
2017 ◽  
Author(s):  
Seema Nandi ◽  
Ashley C. Brown
Keyword(s):  
Cell Migration ◽  
Three Dimensional

scholarly journals Cell migration and organization in three-dimensional in vitro culture driven by stiffness gradient

2016 ◽  
Vol 113 (11) ◽  
pp. 2496-2506 ◽  
Author(s):  
Danielle Joaquin ◽  
Michael Grigola ◽  
Gubeum Kwon ◽  
Christopher Blasius ◽  
Yutao Han ◽  
...  
Keyword(s):  
Cell Migration ◽  
In Vitro Culture ◽  
Three Dimensional

scholarly journals Modeling Liver Organogenesis by Recreating Three-Dimensional Collective Cell Migration: A Role for TGFβ Pathway

2021 ◽  
Vol 9 ◽  
Author(s):  
Ogechi Ogoke ◽  
Osama Yousef ◽  
Cortney Ott ◽  
Allison Kalinousky ◽  
Wayne Lin ◽  
...  
Keyword(s):  
Cell Migration ◽  
Liver Cell ◽  
Transforming Growth Factor Beta ◽  
Transforming Growth Factor ◽  
Three Dimensional ◽  
3D Models ◽  
Branching Morphogenesis ◽  
Lung Fibroblasts ◽  
Collective Cell Migration

Three-dimensional (3D) collective cell migration (CCM) is critical for improving liver cell therapies, eliciting mechanisms of liver disease, and modeling human liver development and organogenesis. Mechanisms of CCM differ in 2D vs. 3D systems, and existing models are limited to 2D or transwell-based systems, suggesting there is a need for improved 3D models of CCM. To recreate liver 3D CCM, we engineered in vitro 3D models based upon a morphogenetic transition that occurs during liver organogenesis, which occurs rapidly between E8.5 and E9.5 (mouse). During this morphogenetic transition, 3D CCM exhibits co-migration (multiple cell types), thick-strand interactions with surrounding septum transversum mesenchyme (STM), branching morphogenesis, and 3D interstitial migration. Here, we engineer several 3D in vitro culture systems, each of which mimics one of these processes in vitro. In mixed spheroids bearing both liver cells and uniquely MRC-5 (fetal lung) fibroblasts, we observed evidence of co-migration, and a significant increase in length and number of liver spheroid protrusions, which was highly sensitive to transforming growth factor beta 1 (TGFβ1) stimulation. In MRC-5-conditioned medium (M-CM) experiments, we observed dose-dependent branching morphogenesis associated with an upregulation of Twist1, which was inhibited by a broad TGFβ inhibitor. In models in which liver spheroids and MRC-5 spheroids were co-cultured, we observed complex strand morphogenesis, whereby thin, linear, 3D liver cell strands attach to the MRC-5 spheroid, anchor and thicken to form permanent and thick anchoring contacts between the two spheroids. In these spheroid co-culture models, we also observed spheroid fusion and strong evidence for interstitial migration. In conclusion, we present several novel cultivation systems that recreate distinct features of liver 3D CCM. These methodologies will greatly improve our molecular, cellular, and tissue-scale understanding of liver organogenesis, liver diseases like cancer, and liver cell therapy, and will also serve as a tool to bridge conventional 2D studies and preclinical in vivo studies.

scholarly journals Reduction of Human Glioblastoma Spheroids Using Cold Atmospheric Plasma: The Combined Effect of Short- and Long-Lived Reactive Species

Cancers ◽  
2018 ◽  
Vol 10 (11) ◽  
pp. 394 ◽  
Author(s):  
Angela Privat-Maldonado ◽  
Yury Gorbanev ◽  
Sylvia Dewilde ◽  
Evelien Smits ◽  
Annemie Bogaerts
Keyword(s):  
Cell Migration ◽  
Tumour Size ◽  
Three Dimensional ◽  
Tumour Microenvironment ◽  
Atmospheric Plasma ◽  
Paramagnetic Resonance ◽  
Cold Atmospheric Plasma ◽  
And Control ◽  
Phosphate Buffered Saline

Cold atmospheric plasma (CAP) is a promising technology against multiple types of cancer. However, the current findings on the effect of CAP on two-dimensional glioblastoma cultures do not consider the role of the tumour microenvironment. The aim of this study was to determine the ability of CAP to reduce and control glioblastoma spheroid tumours in vitro. Three-dimensional glioblastoma spheroid tumours (U87-Red, U251-Red) were consecutively treated directly and indirectly with a CAP using dry He, He + 5% H2O or He + 20% H2O. The cytotoxicity and spheroid shrinkage were monitored using live imaging. The reactive oxygen and nitrogen species produced in phosphate buffered saline (PBS) were measured by electron paramagnetic resonance (EPR) and colourimetry. Cell migration was also assessed. Our results demonstrate that consecutive CAP treatments (He + 20% H2O) substantially shrank U87-Red spheroids and to a lesser degree, U251-Red spheroids. The cytotoxic effect was due to the short- and long-lived species delivered by CAP: they inhibited spheroid growth, reduced cell migration and decreased proliferation in CAP-treated spheroids. Direct treatments were more effective than indirect treatments, suggesting the importance of CAP-generated, short-lived species for the growth inhibition and cell cytotoxicity of solid glioblastoma tumours. We concluded that CAP treatment can effectively reduce glioblastoma tumour size and restrict cell migration, thus demonstrating the potential of CAP therapies for glioblastoma.

scholarly journals Mesenchyme modulates three-dimensional, collective cell migration of liver cells in vitro - a role for TGFß pathway

2020 ◽  
Author(s):  
Ogechi Ogoke ◽  
Osama Yousef ◽  
Cortney Ott ◽  
Allison Kalinousky ◽  
Lin Wayne ◽  
...  
Keyword(s):  
Cell Migration ◽  
Liver Cell ◽  
Three Dimensional ◽  
Branching Morphogenesis ◽  
Lung Fibroblasts ◽  
Collective Cell Migration ◽  
Paracrine Effects ◽  
Cell Therapies

ABSTRACTThree dimensional (3D) collective cell migration (CCM) is critical for improving liver cell therapies, eliciting mechanisms of liver disease, and modeling human liver development/ organogenesis. Here, we modeled liver organogenesis to induce 3D CCM and improve existing models. The liver diverticulum, normally surrounded by septum transversum mesenchyme (STM) at E8.5, was modeled with a miniature liver spheroid surrounded by mesenchymal cells and matrix. In mixed spheroid models with both liver and uniquely MRC5 (fetal lung) fibroblasts, we observed co-migration of cells, and a significant increase in length and number of liver spheroid protrusions, and this was highly sensitive to TGFB1 stimulation. To understand paracrine effects between MRC-5 cells and liver, we performed conditioned medium (M-CM) experiments. Interestingly, the addition of M-CM increased liver 3D CCM, with thin, 3D, dose-dependent branching morphogenesis, an upregulation of Twist1, and a sensitivity to a broad TGFB inhibitor. To test the effects of cell-cell interactions of 3D CCM, the STM was modeled with a spheroid of MRC-5 cells, and we performed co-spheroid culture of liver with MRC-5. We observed a complex morphogenesis, whereby thin, linear, 3D liver cell strands attach to the MRC-5 spheroid, anchor, and thicken to form permanent and thick anchoring contacts between the two spheroids. We also observed spheroid fusion, a form of interstitial migration. In conclusion, we present several novel cultivation systems that induce distinct features of 3D CCM, as judged by the presence of branching, linearity, thickness, and interstitial migration. These methodologies will greatly improve our molecular, cellular, and tissue-scale understanding of liver organogenesis, liver diseases, and liver cell therapy, and will serve as a tool to bridge conventional 2D studies and preclinical in vivo studies.

scholarly journals Development of Gelatin-Based Flexible Three-Dimensional Capillary Pattern Microfabrication Technology for Analysis of Collective Cell Migration

2021 ◽  
Vol 4 (1) ◽  
pp. 11
Author(s):  
Hiromichi Hashimoto ◽  
Mitsuru Sentoku ◽  
Kento Iida ◽  
Kenji Yasuda
Keyword(s):  
Cell Migration ◽  
Structural Changes ◽  
Vascular Endothelial Cells ◽  
Three Dimensional ◽  
Migration Velocity ◽  
In Vitro Assays ◽  
Collective Cell Migration ◽  
Interactive Behavior ◽  
Microfabrication Technology

Collective cell migration is thought to be a dynamic and interactive behavior of cell cohorts that is essential for diverse physiological developments in living organisms. Recent studies revealed that the topographical properties of the environment regulate the migration modes of cell cohorts, such as diffusion versus contraction relaxation transport and the appearance of vortices in larger available space. However, conventional in vitro assays fail to observe changes in cell behavior in response to the structural changes. In this study, we developed a method to fabricate the flexible three-dimensional structures of capillary microtunnels to examine the behavior of vascular endothelial cells (ECs). Microtunnels with altering diameters were formed inside gelatin gel through spot heating a portion of gelatin by irradiating the µm-sized absorption at the tip of the microneedle with a focused permeable 1064 nm infrared laser. The ECs moved and spread two-dimensionally on the inner surface of the capillary microtunnels as a monolayer instead of filling the capillary. In contrast to the 3D straight topographical constraint, which exhibited width-dependent migration velocity, the leading ECs altered its migration velocity according to the change in the supply of cells behind the leading ECs, caused by their progression through the diameter-altering structure. Our findings provide insights into the collective migration modes inside 3D confinement structures, including their fluid-like behavior and the conservation of cell numbers.

Digital holographic microscopy for the three-dimensional dynamic analysis of in vitro cancer cell migration

2006 ◽  
Vol 11 (5) ◽  
pp. 054032 ◽  
Author(s):  
Frank Dubois ◽  
Catherine Yourassowsky ◽  
Olivier Monnom ◽  
Jean-Claude Legros ◽  
Olivier Debeir ◽  
...  
Keyword(s):  
Cell Migration ◽  
Dynamic Analysis ◽  
Cancer Cell ◽  
Three Dimensional ◽  
Digital Holographic Microscopy ◽  
Cancer Cell Migration ◽  
Holographic Microscopy
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