Rotary Culture Enhances Pre-osteoblast Aggregation and Mineralization

2005 ◽  
Vol 84 (6) ◽  
pp. 542-547 ◽  
Author(s):  
S.R. Facer ◽  
R.S. Zaharias ◽  
M.E. Andracki ◽  
J. Lafoon ◽  
S.K. Hunter ◽  
...  
Keyword(s):  
Three Dimensional ◽  
Cell Aggregation ◽  
Osteogenic Potential ◽  
Tissue Culture Plastic ◽  
Osteoblast Cells ◽  
3D Cultures ◽  
Cell Culture Systems ◽  
3D Environments ◽  
Rotary Culture ◽  
Potential Use

Three-dimensional environments have been shown to enhance cell aggregation and osteoblast differentiation. Thus, we hypothesized that three-dimensional (3D) growth environments would enhance the mineralization rate of human embryonic palatal mesenchymal (HEPM) pre-osteoblasts. The objective of this study was to investigate the potential use of rotary cell culture systems (RCCS) as a means to enhance the osteogenic potential of pre-osteoblast cells. HEPM cells were cultured in a RCCS to create 3D enviroments. Tissue culture plastic (2D) cultures served as our control. 3D environments promoted three-dimensional aggregate formations. Increased calcium and phosphorus deposition was significantly enhanced three- to 18-fold (P < 0.001) in 3D cultures as compared with 2D environments. 3D cultures mineralized in 1 wk as compared with the 2D cultures, which took 4 wks, a decrease in time of nearly 75%. In conclusion, our studies demonstrated that 3D environments enhanced osteoblast cell aggregation and mineralization.

scholarly journals Fibronectin Matrix Assembly Regulates α5β1-mediated Cell Cohesion

2004 ◽  
Vol 15 (3) ◽  
pp. 973-981 ◽  
Author(s):  
Elizabeth E. Robinson ◽  
Ramsey A. Foty ◽  
Siobhan A. Corbett
Keyword(s):  
Three Dimensional ◽  
Cell Aggregation ◽  
Alternative Mechanism ◽  
Matrix Assembly ◽  
Integrin Α4 ◽  
3D Environments ◽  
Cellular Aggregates ◽  
Α5β1 Integrin ◽  
Cells Cultured

Integrin-extracellular matrix (ECM) interactions in two-dimensional (2D) culture systems are widely studied (Goldstein and DiMilla, 2002. J Biomed. Mater. Res. 59, 665–675; Koo et al., 2002. J. Cell Sci. 115, 1423–1433). Less understood is the role of the ECM in promoting intercellular cohesion in three-dimensional (3D) environments. We have demonstrated that the α5β1-integrin mediates strong intercellular cohesion of 3D cellular aggregates (Robinson et al., 2003. J. Cell Sci. 116, 377–386). To further investigate the mechanism of α5β1-mediated cohesivity, we used a series of chimeric α5β1-integrin–expressing cells cultured as multilayer cellular aggregates. In these cell lines, the α5 subunit cytoplasmic domain distal to the GFFKR sequence was truncated, replaced with that of the integrin α4, the integrin α2, or maintained intact. Using these cells, α5β1-integrin–mediated cell aggregation, compaction and cohesion were determined and correlated with FN matrix assembly. The data presented demonstrate that cells cultured in the absence of external mechanical support can assemble a FN matrix that promotes integrin-mediated aggregate compaction and cohesion. Further, inhibition of FN matrix assembly blocks the intercellular associations required for compaction, resulting in cell dispersal. These results demonstrate that FN matrix assembly contributes significantly to tissue cohesion and represents an alternative mechanism for regulating tissue architecture.

scholarly journals Mammary gland 3D cell culture systems in farm animals

2021 ◽  
Vol 52 (1) ◽  
Author(s):  
Laurence Finot ◽  
Eric Chanat ◽  
Frederic Dessauge
Keyword(s):  
Cell Culture ◽  
Mammary Gland ◽  
Bacterial Infections ◽  
Three Dimensional ◽  
3D Cell Culture ◽  
Farm Animals ◽  
Culture Systems ◽  
Cell Culture Systems

AbstractIn vivo study of tissue or organ biology in mammals is very complex and progress is slowed by poor accessibility of samples and ethical concerns. Fortunately, however, advances in stem cell identification and culture have made it possible to derive in vitro 3D “tissues” called organoids, these three-dimensional structures partly or fully mimicking the in vivo functioning of organs. The mammary gland produces milk, the source of nutrition for newborn mammals. Milk is synthesized and secreted by the differentiated polarized mammary epithelial cells of the gland. Reconstructing in vitro a mammary-like structure mimicking the functional tissue represents a major challenge in mammary gland biology, especially for farm animals for which specific agronomic questions arise. This would greatly facilitate the study of mammary gland development, milk secretion processes and pathological effects of viral or bacterial infections at the cellular level, all with the objective of improving milk production at the animal level. With this aim, various 3D cell culture models have been developed such as mammospheres and, more recently, efforts to develop organoids in vitro have been considerable. Researchers are now starting to draw inspiration from other fields, such as bioengineering, to generate organoids that would be more physiologically relevant. In this chapter, we will discuss 3D cell culture systems as organoids and their relevance for agronomic research.

Cell-to-cell contact influences proliferative marker expression and apoptosis in MIN6 cells grown in islet-like structures

2005 ◽  
Vol 288 (3) ◽  
pp. E502-E509 ◽  
Author(s):  
Melanie J. Luther ◽  
Emma Davies ◽  
Dany Muller ◽  
Moira Harrison ◽  
Adrian J. Bone ◽  
...  
Keyword(s):  
Cellular Proliferation ◽  
Kinase Inhibitors ◽  
Three Dimensional ◽  
Cell Aggregation ◽  
Cell Interactions ◽  
Cell Phenotype ◽  
Cell Contact ◽  
Min6 Cells ◽  
Primary Mouse ◽  
E Cadherin

Cell-to-cell interactions play an important role in the development and maintenance of the β-cell phenotype. Here, we have investigated whether E-cadherin plays a role in regulating the growth of insulin-secreting MIN6 cells configured as three-dimensional islet-like clusters (pseudoislets). Pseudoislets form by cell aggregation rather than by proliferation from individual cells and attain the size of primary mouse islets after ∼7 days of maintenance in culture. E-cadherin is known to mediate homotypic cell adhesion between β-cells and has also been implicated in a number of cellular processes, including proliferation, apoptosis, and differentiation. E-cadherin and its associated intracellular elements, α- and β-catenin, were upregulated in MIN6 pseudoislets. Pseudoislet formation was associated with an increased expression of cyclin-dependent kinase inhibitors and a concomitant downregulation of Ki67, suggesting an overall reduction in cellular proliferation. However, measurements of 5-bromo-2′-deoxyuridine incorporation revealed that there were no differences in the rate of MIN6 cell proliferation whether they were configured as monolayers or as pseudoislets, which is likely to be a result of their being a transformed cell line. Cells within pseudoislets were not necrotic, but apoptosis appeared to be upregulated in the islet-like structures. However, no differential expression of Fas and FasL was detected in monolayers and pseudoislets. These results suggest that cell-to-cell interactions within islet-like structures may initiate antiproliferative and proapoptotic signals.

The influence of proepicardial cells on the osteogenic potential of marrow stromal cells in a three-dimensional tubular scaffold

Biomaterials ◽  
2008 ◽  
Vol 29 (14) ◽  
pp. 2203-2216 ◽  
Author(s):  
Mani T. Valarmathi ◽  
Michael J. Yost ◽  
Richard L. Goodwin ◽  
Jay D. Potts
Keyword(s):  
Stromal Cells ◽  
Three Dimensional ◽  
Marrow Stromal Cells ◽  
Osteogenic Potential ◽  
Tubular Scaffold

scholarly journals Application of microfluidic systems for neural differentiation of cells

2019 ◽  
Vol 2 (4) ◽  
pp. 370-381
Author(s):  
Zahra Hesari ◽  
Fatemeh Mottaghitalab ◽  
Akram Shafiee ◽  
Masoud Soleymani ◽  
Rasoul Dinarvand ◽  
...  
Keyword(s):  
Stem Cells ◽  
Small Molecules ◽  
Neuronal Differentiation ◽  
Neural Differentiation ◽  
Three Dimensional ◽  
Microfluidic System ◽  
Microfluidic Systems ◽  
Cell Culture Systems ◽  
Novel Model ◽  
Dimensional Cell

Neural differentiation of stem cells is an important issue in development of central nervous system. Different methods such as chemical stimulation with small molecules, scaffolds, and microRNA can be used for inducing the differentiation of neural stem cells. However, microfluidic systems with the potential to induce neuronal differentiation have established their reputation in the field of regenerative medicine. Organization of microfluidic system represents a novel model that mimic the physiologic microenvironment of cells among other two and three dimensional cell culture systems. Microfluidic system has patterned and well-organized structure that can be combined with other differentiation techniques to provide optimal conditions for neuronal differentiation of stem cells. In this review, different methods for effective differentiation of stem cells to neuronal cells are summarized. The efficacy of microfluidic systems in promoting neuronal differentiation is also addressed.

scholarly journals Evaluation of RGD functionalization in hybrid hydrogels as 3D neural stem cell culture systems

2018 ◽  
Vol 6 (3) ◽  
pp. 501-510 ◽  
Author(s):  
Emanuele Mauri ◽  
Alessandro Sacchetti ◽  
Nunzio Vicario ◽  
Luca Peruzzotti-Jametti ◽  
Filippo Rossi ◽  
...  
Keyword(s):  
Central Nervous System ◽  
Cell Culture ◽  
Culture System ◽  
Three Dimensional ◽  
Cell Culture System ◽  
Nervous System Diseases ◽  
Central Nervous System Diseases ◽  
Hybrid Hydrogels ◽  
Cell Culture Systems ◽  
Dimensional Cell

The use of neural stem cells (NSCs) in cell therapy has become a powerful tool used for the treatment of central nervous system diseases and the design of a three-dimensional cell culture system to improve NSCs viability is a challenge.

An Integrated Platform for Educational Virtual Environments

2008 ◽  
pp. 530-554
Author(s):  
Christos Bouras ◽  
Eleftheria Giannaka ◽  
Maria Nani ◽  
Alexandros Panagopoulos ◽  
Thrasyvoulos Tsiatosos
Keyword(s):  
Virtual Environments ◽  
Online Courses ◽  
Three Dimensional ◽  
Content Management ◽  
Fully Integrated ◽  
3D Space ◽  
Learning Content ◽  
Asynchronous Courses ◽  
Integrated Platform ◽  
3D Environments

In this chapter, we present the design and implementation of an integrated platform for Educational Virtual Environments. This platform aims to support an educational community, synchronous online courses in multi-user three-dimensional (3D) environments, and the creation and access of asynchronous courses through a learning content management system. In order to offer synchronous courses, we have implementeda system called EVE-II, which supports stable event sharing for multi-user 3D places, easy creation of multi-user 3D places, H.323-based voice- over IP services fully integrated in a 3D space, as well as many concurrent 3D multi-user spaces.

scholarly journals A 3D Cell Death Assay to Quantitatively Determine Ferroptosis in Spheroids

Cells ◽  
2020 ◽  
Vol 9 (3) ◽  
pp. 703 ◽  
Author(s):  
Robin Demuynck ◽  
Iuliia Efimova ◽  
Abraham Lin ◽  
Heidi Declercq ◽  
Dmitri V. Krysko
Keyword(s):  
Cell Death ◽  
Three Dimensional ◽  
Drug Efficacy ◽  
Cost Effective ◽  
3D Cultures ◽  
Cell Death Assay ◽  
Tumour Spheroids ◽  
A Cell ◽  
Triton X

The failure of drug efficacy in clinical trials remains a big issue in cancer research. This is largely due to the limitations of two-dimensional (2D) cell cultures, the most used tool in drug screening. Nowadays, three-dimensional (3D) cultures, including spheroids, are acknowledged to be a better model of the in vivo environment, but detailed cell death assays for 3D cultures (including those for ferroptosis) are scarce. In this work, we show that a new cell death analysis method, named 3D Cell Death Assay (3DELTA), can efficiently determine different cell death types including ferroptosis and quantitatively assess cell death in tumour spheroids. Our method uses Sytox dyes as a cell death marker and Triton X-100, which efficiently permeabilizes all cells in spheroids, was used to establish 100% cell death. After optimization of Sytox concentration, Triton X-100 concentration and timing, we showed that the 3DELTA method was able to detect signals from all cells without the need to disaggregate spheroids. Moreover, in this work we demonstrated that 2D experiments cannot be extrapolated to 3D cultures as 3D cultures are less sensitive to cell death induction. In conclusion, 3DELTA is a more cost-effective way to identify and measure cell death type in 3D cultures, including spheroids.

scholarly journals Exposure of the SH-SY5Y Human Neuroblastoma Cells to 50-Hz Magnetic Field: Comparison Between Two-Dimensional (2D) and Three-Dimensional (3D) In Vitro Cultures

2020 ◽  
Author(s):  
Claudia Consales ◽  
Alessio Butera ◽  
Caterina Merla ◽  
Emanuela Pasquali ◽  
Vanni Lopresto ◽  
...  
Keyword(s):  
Magnetic Field ◽  
Neuroblastoma Cells ◽  
Three Dimensional ◽  
3D Culture ◽  
Cell Cycle Distribution ◽  
Two Dimensional ◽  
3D Scaffold ◽  
Human Neuroblastoma Cells ◽  
Human Neuroblastoma ◽  
3D Cultures

AbstractWe here characterize the response to the extremely low-frequency (ELF) magnetic field (MF, 50 Hz, 1 mT) of SH-SY5Y human neuroblastoma cells, cultured in a three-dimensional (3D) Alvetex® scaffold compared to conventional two-dimensional (2D) monolayers. We proved that the growing phenotype of proliferating SH-SY5Y cells is not affected by the culturing conditions, as morphology, cell cycle distribution, proliferation/differentiation gene expression of 3D-cultures overlap what reported in 2D plates. In response to 72-h exposure to 50-Hz MF, we demonstrated that no proliferation change and apoptosis activation occur in both 2D and 3D cultures. Consistently, no modulation of Ki67, MYCN, CCDN1, and Nestin, of invasiveness and neo-angiogenesis-controlling genes (HIF-1α, VEGF, and PDGF) and of microRNA epigenetic signature (miR-21-5p, miR-222-3p and miR-133b) is driven by ELF exposure. Conversely, intracellular glutathione content and SOD1 expression are exclusively impaired in 3D-culture cells in response to the MF, whereas no change of such redox modulators is observed in SH-SY5Y cells if grown on 2D monolayers. Moreover, ELF-MF synergizes with the differentiating agents to stimulate neuroblastoma differentiation into a dopaminergic (DA) phenotype in the 3D-scaffold culture only, as growth arrest and induction of p21, TH, DAT, and GAP43 are reported in ELF-exposed SH-SY5Y cells exclusively if grown on 3D scaffolds. As overall, our findings prove that 3D culture is a more reliable experimental model for studying SH-SY5Y response to ELF-MF if compared to 2D conventional monolayer, and put the bases for promoting 3D systems in future studies addressing the interaction between electromagnetic fields and biological systems.

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