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 Fucoxanthin Holds Potential to Become a Drug Adjuvant in Breast Cancer Treatment: Evidence from 2D and 3D Cell Cultures

Molecules ◽  
2021 ◽  
Vol 26 (14) ◽  
pp. 4288
Author(s):  
Fernanda Malhão ◽  
Ana Catarina Macedo ◽  
Carla Costa ◽  
Eduardo Rocha ◽  
Alice Abreu Ramos
Keyword(s):  
Breast Cancer ◽  
Cell Lines ◽  
3D Models ◽  
Anticancer Activities ◽  
Cytotoxic Effects ◽  
3D Cultures ◽  
3D Cell Cultures ◽  
Tumoral Cell ◽  
Microscopy Techniques ◽  
2D And 3D

Fucoxanthin (Fx) is a carotenoid derived from marine organisms that exhibits anticancer activities. However, its role as a potential drug adjuvant in breast cancer (BC) treatment is still poorly explored. Firstly, this study investigated the cytotoxic effects of Fx alone and combined with doxorubicin (Dox) and cisplatin (Cis) on a panel of 2D-cultured BC cell lines (MCF7, SKBR3 and MDA-MB-231) and one non-tumoral cell line (MCF12A). Fucoxanthin induced cytotoxicity against all the cell lines and potentiated Dox cytotoxic effects towards the SKBR3 and MDA-MB-231 cells. The combination triggering the highest cytotoxicity (Fx 10 µM + Dox 1 µM in MDA-MB-231) additionally showed significant induction of cell death and genotoxic effects, relative to control. In sequence, the same combination was tested on 3D cultures using a multi-endpoint approach involving bioactivity assays and microscopy techniques. Similar to 2D cultures, the combination of Fx and Dox showed higher cytotoxic effects on 3D cultures compared to the isolated compounds. Furthermore, this combination increased the number of apoptotic cells, decreased cell proliferation, and caused structural and ultrastructural damages on the 3D models. Overall, our findings suggest Fx has potential to become an adjuvant for Dox chemotherapy regimens in BC treatment.

Extracellular vesicles from three dimensional culture of human placental mesenchymal stem cells ameliorated renal ischemia/reperfusion injury

2021 ◽  
pp. 039139882098680
Author(s):  
Xuefeng Zhang ◽  
Nan Wang ◽  
Yuhua Huang ◽  
Yan Li ◽  
Gang Li ◽  
...  
Keyword(s):  
Stem Cells ◽  
Mesenchymal Stem Cells ◽  
Extracellular Vesicles ◽  
Therapeutic Potential ◽  
Expression Profiles ◽  
Ischemia Reperfusion ◽  
Three Dimensional ◽  
3D Culture ◽  
Placental Mesenchymal Stem Cells ◽  
2D And 3D

Background: Three-dimensional (3D) culture has been reported to increase the therapeutic potential of mesenchymal stem cells (MSCs). The present study assessed the therapeutic efficacy of extracellular vesicles (EVs) from 3D cultures of human placental MSCs (hPMSCs) for acute kidney injury (AKI). Methods: The supernatants from monolayer culture (2D) and 3D culture of hPMSCs were ultra-centrifuged for EVs isolation. C57BL/6 male mice were submitted to 45 min bilateral ischemia of kidney, followed by renal intra-capsular administration of EVs within a 72 h reperfusion period. Histological, immunohistochemical, and ELISA analyses of kidney samples were performed to evaluate cell death and inflammation. Kidney function was evaluated by measuring serum creatinine and urea nitrogen. The miRNA expression profiles of EVs from 2D and 3D culture of hPMSCs were evaluated using miRNA microarray analysis. Results: The 3D culture of hPMSCs formed spheroids with different diameters depending on the cell density seeded. The hPMSCs produced significantly more EVs in 3D culture than in 2D culture. More importantly, injection of EVs from 3D culture of hPMSCs into mouse kidney with ischemia-reperfusion (I/R)-AKI was more beneficial in protecting from progression of I/R than those from 2D culture. The EVs from 3D culture of hPMSCs were more efficient against apoptosis and inflammation than those from 2D culture, which resulted in a reduction in tissue damage and amelioration of renal function. MicroRNA profiling analysis revealed that a set of microRNAs were significantly changed in EVs from 3D culture of hPMSCs, especially miR-93-5p. Conclusion: The EVs from 3D culture of hPMSCs have therapeutic potential for I/R-AKI.

scholarly journals 3D Culture Systems for Exploring Cancer Immunology

Cancers ◽  
2020 ◽  
Vol 13 (1) ◽  
pp. 56
Author(s):  
Allison A. Fitzgerald ◽  
Eric Li ◽  
Louis M. Weiner
Keyword(s):  
Immune Cell ◽  
Three Dimensional ◽  
3D Culture ◽  
3D Models ◽  
Therapeutic Benefit ◽  
Cancer Immunology ◽  
Microfluidic Chips ◽  
Murine Models ◽  
Human Immune System ◽  
Culture Systems

Cancer immunotherapy has revolutionized cancer treatment, spurring extensive investigation into cancer immunology and how to exploit this biology for therapeutic benefit. Current methods to investigate cancer-immune cell interactions and develop novel drug therapies rely on either two-dimensional (2D) culture systems or murine models. However, three-dimensional (3D) culture systems provide a potentially superior alternative model to both 2D and murine approaches. As opposed to 2D models, 3D models are more physiologically relevant and better replicate tumor complexities. Compared to murine models, 3D models are cheaper, faster, and can study the human immune system. In this review, we discuss the most common 3D culture systems—spheroids, organoids, and microfluidic chips—and detail how these systems have advanced our understanding of cancer immunology.

Enhanced growth and myogenic differentiation of spheroid-derived C2C12 cells

2021 ◽  
Author(s):  
Guang-Zhen Jin
Keyword(s):  
Myogenic Differentiation ◽  
Three Dimensional ◽  
3D Culture ◽  
Stem Cell Differentiation ◽  
Culture Conditions ◽  
C2c12 Cells ◽  
Dependent Manner ◽  
Physical Force ◽  
Myod Gene ◽  
2D And 3D

Abstract Among many factors of controlling stem cell differentiation, the key transcription factor upregulation via physical force is a good strategy on the lineage-specific differentiation of stem cells. The study aimed to compare growth and myogenic potentials between the parental cells (PCs) and the 1-day-old C2C12 spheroid-derived cells (SDCs) in two-dimensional (2D) and three-dimensional (3D) culture conditions through examination of the cell proliferation and the expression of myogenic genes. The data showed that 1-day-old spheroids had more intense expression of MyoD gene with respect to the PCs. The proliferation of the SDCs significantly higher than the PCs in a time dependent manner. The SDCs had also significantly higher myogenic potential than the PCs in 2D and 3D culture conditions. The results suggest that MyoD gene upregulation through cell-cell contacts is the good approach for preparation of seed cells in muscle tissue engineering.

scholarly journals Inhibitors of A Disintegrin And Metalloproteinases-10 reduce Hodgkin lymphoma cell growth in 3D microenvironments and enhance brentuximab-vedotin effect

Haematologica ◽  
2021 ◽  
Author(s):  
Roberta Pece ◽  
Sara Tavella ◽  
Delfina Costa ◽  
Serena Varesano ◽  
Caterina Camodeca ◽  
...  
Keyword(s):  
Hodgkin Lymphoma ◽  
Drug Testing ◽  
Cell Damage ◽  
Three Dimensional ◽  
3D Culture ◽  
Glucose Consumption ◽  
3D Models ◽  
Brentuximab Vedotin ◽  
Collagen Scaffolds ◽  
Antibody Drug Conjugate

Shedding of A Disintegrin And Metalloproteinases (ADAM10) substrates, like TNFα or CD30, can affect both anti-tumor immune response and antibody-drug-conjugate (ADC)-based immunotherapy. We have published two new ADAM10 inhibitors, LT4 and MN8 able to prevent such shedding in Hodgkin lymphoma (HL). Since tumor tissue architecture deeply influence the outcome of anti-cancer treatments, we set up new three-dimensional (3D) culture systemsto verify whether ADAM10 inhibitors can contribute to, or enhance, the anti-lymphoma effects of the ADC brentuximab-vedotin (BtxVed).To recapitulate some aspects of lymphoma structure and architecture, we assembled two 3D culture models: mixed spheroids made of HL lymph node (LN) mesenchymal stromal cells (MSC) and Reed Sternberg/Hodgkin lymphoma cells (HL cells) or collagen scaffolds repopulated with LN-MSC and HL cells. In these 3D systems we found that: 1) the ADAM10 inhibitors LT4 and MN8 reduce ATP content or glucose consumption, related to cell proliferation, increasing lactate dehydrogenase (LDH) release as a cell damage hallmark; 2) these events are paralleled by mixed spheroids size reduction and inhibition of CD30 and TNFα shedding; 3) the effects observed can be reproduced in repopulated HL LN-derived matrix or collagen scaffolds; 4) ADAM10 inhibitors enhance the antilymphoma effect of the anti-CD30 ADC BtxVed both in conventional cultures and in repopulated scaffolds. Thus, we provide evidence for direct and combined anti-lymphoma effect of ADAM10 inhibitors with BtxVed, leading to improvement of ADC effects; this is documented in 3D models recapitulating features of LN microenvironment, that can be proposed as reliable tool for antilymphoma drug testing.

Thermally Induced Apoptosis, Necrosis, and Heat Shock Protein Expression in Three-Dimensional Culture

2014 ◽  
Vol 136 (7) ◽  
Author(s):  
Alfred S. Song ◽  
Amer M. Najjar ◽  
Kenneth R. Diller
Keyword(s):  
Prostate Cancer ◽  
Heat Shock ◽  
Heat Shock Protein ◽  
Fluorescent Protein ◽  
Three Dimensional ◽  
3D Culture ◽  
Hsp70 Expression ◽  
Three Dimensional Culture ◽  
Apoptosis And Necrosis ◽  
2D And 3D

This study was conducted to compare the heat shock responses of cells grown in 2D and 3D culture environments as indicated by the level of heat shock protein 70 expression and the incidence of apoptosis and necrosis of prostate cancer cell lines in response to graded hyperthermia. PC3 cells were stably transduced with a dual reporter system composed of two tandem expression cassettes—a conditional heat shock protein promoter driving the expression of green fluorescent protein (HSPp-GFP) and a cytomegalovirus (CMV) promoter controlling the constitutive expression of a “beacon” red fluorescent protein (CMVp-RFP). Two-dimensional and three-dimensional cultures of PC3 prostate cancer cells were grown in 96-well plates for evaluation of their time-dependent response to supraphysiological temperature. To induce controlled hyperthermia, culture plates were placed on a flat copper surface of a circulating water manifold that maintained the specimens within ±0.1 °C of a target temperature. Hyperthermia protocols included various combinations of temperature, ranging from 37 °C to 57 °C, and exposure times of up to 2 h. The majority of protocols were focused on temperature and time permutations, where the response gradient was greatest. Post-treatment analysis by flow cytometry analysis was used to measure the incidences of apoptosis (annexin V-FITC stain), necrosis (propidium iodide (PI) stain), and HSP70 transcription (GFP expression). Cells grown in 3D compared with 2D culture showed reduced incidence of apoptosis and necrosis and a higher level of HSP70 expression in response to heat shock at the temperatures tested. Cells responded differently to hyperthermia when grown in 2D and 3D cultures. Three-dimensional culture appears to enhance survival plausibly by activating protective processes related to enhanced-HSP70 expression. These differences highlight the importance of selecting physiologically relevant 3D models in assessing cellular responses to hyperthermia in experimental settings.

Effects of Weld Geometry Size on the Pressure-Bearing Capacity of an API X80 LSAW Pipe

2020 ◽  
Vol 142 (5) ◽  
Author(s):  
Gui-ying Qiao ◽  
Yu-meng Liu ◽  
Jun-si Wang ◽  
Fu-ren Xiao
Keyword(s):  
Stress Distribution ◽  
Weld Joint ◽  
Large Diameter ◽  
Three Dimensional ◽  
3D Models ◽  
Welding Parameter ◽  
Weld Geometry ◽  
Transmission Pipeline ◽  
2D And 3D ◽  
Submerged Arc

Abstract The weld joint is the weakest zone of a longitudinal-seam submerged arc welded (LSAW) pipe, which has great effects on its in-service properties and safety. The weld geometry and shape of the weld joint are important factors that affect the mechanical properties of the pipe. In this work, two-dimensional (2D) and three-dimensional (3D) finite element models (FEMs) of a large-diameter, heavy-wall API X80 LSAW pipe were established, and the stress distribution of the pipe was calculated to simulate the in-service gas transmission pipeline. Results showed that the stress distribution calculated by both 2D and 3D models are similar. Consequently, the effects of weld geometry, softening of heat affected zone (HAZ), and strength matching of the weld joint on bearing pressure capability of the pipe were analyzed, and the results showed that the bearing pressure capability of the pipe can be improved by controlling these parameters. These results are beneficial in providing references for welding parameter design and improved properties of the X80 LSAW pipe.

Development of Three-Dimensional DC Electric Field Stimulation Bio-Reactor for Axonal Outgrowth Enhancement

2018 ◽  
Author(s):  
Shohei Tanaka ◽  
Ryota Sakiyama ◽  
Koji Yamamoto ◽  
Yusuke Morita ◽  
Eiji Nakamachi
Keyword(s):  
Electric Field ◽  
Three Dimensional ◽  
3D Culture ◽  
Collagen Gel ◽  
Control Group ◽  
Axonal Outgrowth ◽  
The Neural Network ◽  
Dc Electric Field ◽  
Current Electric Field

Numerous studies of electrical stimulation effects on the nerve regeneration have been carried out. However, there were very few investigations which adopt the 3D culture that mimics the in vivo environment. In this study, we designed and fabricated a new 3D direct current electric field (DCEF) stimulation bio-reactor and investigated the effectiveness on the axonal outgrowth enhancement. We searched an optimum structure using the finite element (FE) analyses to obtain a uniform DCEF in the culture region. A measurement result of DCEF strength showed an agreement with FE results. The rat phenocromocytoma cells (PC12) were disseminated in the collagen gel and 3D culture was performed. We observed the morphologies of cell bodies and neurites using the multiphoton excitation fluorescence microscope (MPM). Both increases in 11.3% of mean axonal length and in 4.2% of axogenesis rate, under the condition of 5.0 mV/mm on 6 hours a day for 4 days, were obtained. Further, there was a tendency of longer connecting distance between cell bodies in the DCEF group than one in the Control group. As a result, we validated the efficacies of our stimulation, both for the axonal extension and the neural network generation, using our newly developed bio-reactor.

Linear and Nonlinear Model of Cutting Forces in Peripheral Milling: A Comparison Between 2D and 3D Models

2010 ◽  
Author(s):  
Hamed Moradi ◽  
Mohammad R. Movahhedy ◽  
Gholamreza Vossoughi
Keyword(s):  
Cutting Forces ◽  
Nonlinear Models ◽  
Three Dimensional ◽  
Chip Thickness ◽  
Maximum Error ◽  
3D Models ◽  
Peripheral Milling ◽  
Third Order ◽  
Order Polynomial ◽  
2D And 3D

Peripheral milling is extensively used in manufacturing processes, especially in aerospace industry where end mills are used for milling of wing parts and engine components. Knowledge of the cutting forces is the first necessary stage in analysis of the milling process. In this paper, cutting forces are presented for both two and three dimensional models. Instead of the common linear dependency of cutting forces to the cut chip thickness, two nonlinear models are presented. In the first model, cutting forces are considered as a function of chip thickness with a complete third order polynomial. In the second one, the quadratic and constant terms of the third order polynomial are set to zero. Results show about 2–3% and 2–7% maximum error between the linear, first and second nonlinear models, for 2D and 3D models, respectively. According to the simulation results, both the 2D and 3D models with second type of nonlinearity can be effectively used in practice. The advantage of such modelling is its simplicity in nonlinear analysis of the problem based on perturbation techniques.

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