scholarly journals Evaluation of the effect of hydrocortisone in 2D and 3D HEp-2 cell culture

2021 ◽  
Author(s):  
Marcelo Oliveira Fonseca ◽  
Bruno Henrique Godoi ◽  
Newton Soares da Silva ◽  
Cristina Pacheco-Soares
Keyword(s):  
Prolonged Exposure ◽  
3D Culture ◽  
Treated Group ◽  
Mitochondrial Activity ◽  
Stress Hormone ◽  
Primary Stress ◽  
Violet Crystal ◽  
Human Stress ◽  
Keywords Stress ◽  
2D And 3D

Cancer is one of the diseases with the highest incidence globally and that associated with the patient's emotional state, can act positively or negatively in the treatment. Cortisol is a principal primary stress hormone in the human body. The corticoids can increase cell proliferation and reactive oxygen species that contribute to DNA damage. Prolonged exposure to stress can contribute to tissues becoming insensitive to cortisol, the primary human stress hormone. This study explores cortisol's influence on tumor cell development, particularly in human cells of carcinoma of the human laryngeal (HEp-2). HEp-2 cells were exposed to increasing cortisol (hydrocortisone) concentrations for 24 or 48 hours, and cytotoxicity (MTT assay) proliferation assay (crystal violet assay), and immunolabeled 3D culture for fibronectin and FAK were analyzed. The group treated with hydrocortisone showed a significant increase in mitochondrial activity, as for the evaluation by the violet crystal, the treated group showed similar behavior to the control. The 3D culture showed dispersed cells within 24 hours with reduced FAK labeling; however, no changes were observed within 48 hours. Although some cases favored corticosteroid use in cancer patients, a more detailed analysis is necessary before prescribing them. Keywords: Stress, carcinoma, 3D culture, hydrocortisone.

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 Direct comparison of 3D and 2D cultivation reveals higher osteogenic capacity of elderly osteoblasts in 3D

2021 ◽  
Vol 16 (1) ◽  
Author(s):  
Stephan Payr ◽  
Elizabeth Rosado-Balmayor ◽  
Thomas Tiefenboeck ◽  
Tim Schuseil ◽  
Marina Unger ◽  
...  
Keyword(s):  
Gene Expression ◽  
Collagen Type ◽  
3D Culture ◽  
Osteogenic Potential ◽  
Donor Age ◽  
Collagen Type 1 ◽  
2D And 3D ◽  
Human Primary Osteoblasts ◽  
Gene Expression Levels

Abstract Background The aim of this study was the investigation of the osteogenic potential of human osteoblasts of advanced donor age in 2D and 3D culture. Methods Osteoblasts were induced to osteogenic differentiation and cultivated, using the same polystyrene material in 2D and 3D culture for 2 weeks. Samples were taken to evaluate alkaline phosphatase (ALP) activity, mineralization and gene expression. Results Osteoprotegerin (OPG) levels were significantly increased (8.2-fold) on day 7 in 3D compared to day 0 (p < 0.0001) and 11.6-fold higher in 3D than in 2D (p < 0.0001). Both culture systems showed reduced osteocalcin (OC) levels (2D 85% and 3D 50% of basic value). Collagen type 1 (Col1) expression was elevated in 3D on day 7 (1.4-fold; p = 0.009). Osteopontin (OP) expression showed 6.5-fold higher levels on day 7 (p = 0.002) in 3D than in 2D. Mineralization was significantly higher in 3D on day 14 (p = 0.0002). Conclusion Advanced donor age human primary osteoblasts reveal significantly higher gene expression levels of OPG, Col1 and OP in 3D than in monolayer. Therefore, it seems that a relatively high potential of bone formation in a natural 3D arrangement is presumably still present in osteoblasts of elderly people. Trial registration 5217/11 on the 22nd of Dec. 2011.

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 Phytoestrogen (Daidzein) Promotes Chondrogenic Phenotype of Human Chondrocytes in 2D and 3D Culture Systems

2017 ◽  
Vol 14 (2) ◽  
pp. 103-112 ◽  
Author(s):  
Suhaeb A. Mahmod ◽  
Simmrat Snigh ◽  
Ivan Djordjevic ◽  
Yong Mei Yee ◽  
Rohana Yusof ◽  
...  
Keyword(s):  
3D Culture ◽  
Human Chondrocytes ◽  
Culture Systems ◽  
Chondrogenic Phenotype ◽  
2D And 3D

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 The role of the secondary stress in teaching the English rhythm

2018 ◽  
Vol 7 (3) ◽  
pp. 101-106
Author(s):  
Metin Yurtbasi
Keyword(s):  
Stress Function ◽  
Secondary Stress ◽  
Function Words ◽  
Intended Meaning ◽  
Primary Stress ◽  
Or Groups ◽  
Keywords Stress

In the phonological literature in English, which is a stress-timed language, the existence of at least three levels of stress is usually taken for granted. Words, phrases, utterances or sentences have a prominent element in one of their syllables, which usually correlates with a partner in the same unit, called the secondary stress. It so happens that in multi-syllable words or groups bearing more than two content words, there is also a tertiary stress. Function words neighbouring the content words are usually not stressed or they are reduced. In standard writing, the primary stress is indicated by an acute accent, the secondary stress by a grave accent while the lack of stress is not marked at all. This article proposes that the distinction between primary and secondary stress should have more prevalence in the pronunciation teaching curricula, as they expose the rhythm of English and convey the intended meaning clearly.   Keywords: Stress-timed language, primary stress, secondary stress, tertiary stress, rhythm.  

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.

scholarly journals Neural cell responses to wear debris from metal-on-metal total disc replacements

2019 ◽  
Vol 29 (11) ◽  
pp. 2701-2712
Author(s):  
H. Lee ◽  
J. B. Phillips ◽  
R. M. Hall ◽  
Joanne L. Tipper
Keyword(s):  
Stainless Steel ◽  
Cell Viability ◽  
Glial Cell ◽  
Wear Debris ◽  
Particle Production ◽  
3D Culture ◽  
Dna Integrity ◽  
Wear Particles ◽  
Astrocyte Activation ◽  
2D And 3D

Purpose Abstract Total disc replacements, comprising all-metal articulations, are compromised by wear and particle production. Metallic wear debris and ions trigger a range of biological responses including inflammation, genotoxicity, cytotoxicity, hypersensitivity and pseudotumour formation, therefore we hypothesise that, due to proximity to the spinal cord, glial cells may be adversely affected. Methods Clinically relevant cobalt chrome (CoCr) and stainless steel (SS) wear particles were generated using a six-station pin-on-plate wear simulator. The effects of metallic particles (0.5–50 μm3 debris per cell) and metal ions on glial cell viability, cellular activity (glial fibrillary acidic protein (GFAP) expression) and DNA integrity were investigated in 2D and 3D culture using live/dead, immunocytochemistry and a comet assay, respectively. Results CoCr wear particles and ions caused significant reductions in glial cell viability in both 2D and 3D culture systems. Stainless steel particles did not affect glial cell viability or astrocyte activation. In contrast, ions released from SS caused significant reductions in glial cell viability, an effect that was especially noticeable when astrocytes were cultured in isolation without microglia. DNA damage was observed in both cell types and with both biomaterials tested. CoCr wear particles had a dose-dependent effect on astrocyte activation, measured through expression of GFAP. Conclusions The results from this study suggest that microglia influence the effects that metal particles have on astrocytes, that SS ions and particles play a role in the adverse effects observed and that SS is a less toxic biomaterial than CoCr alloy for use in spinal devices. Graphic abstract These slides can be retrieved under Electronic Supplementary Material.

Sign in / Sign up

Export Citation Format

Share Document