scholarly journals Decellularized Human Gut as a Natural 3D Platform for Research in Intestinal Fibrosis

2019 ◽  
Vol 25 (11) ◽  
pp. 1740-1750 ◽  
Author(s):  
Paolo Giuffrida ◽  
Marco Curti ◽  
Walid Al-Akkad ◽  
Carin Biel ◽  
Claire Crowley ◽  
...  
Keyword(s):  
Cell Cultures ◽  
Drug Testing ◽  
Biomarker Discovery ◽  
Disease Modeling ◽  
3D Culture ◽  
Protein Composition ◽  
Intestinal Fibrosis ◽  
3D Cell Cultures ◽  
3D Architecture ◽  
Inflammatory Bowel

Abstract Background The current methodologies for the identification of therapeutic targets for inflammatory bowel disease (IBD) are limited to conventional 2-dimensional (2D) cell cultures and animal models. The use of 3D decellularized human intestinal scaffolds obtained from surgically resected intestine and engineered with human intestinal cells may provide a major advancement in the development of innovative intestinal disease models. The aim of the present study was to design and validate a decellularization protocol for the production of acellular 3D extracellular matrix (ECM) scaffolds from the human duodenum. Methods Scaffolds were characterized by verifying the preservation of the ECM protein composition and 3D architecture of the native intestine and were employed for tissue engineering with primary human intestinal myofibroblasts for up to 14 days. Results Engrafted cells showed the ability to grow and remodel the surrounding ECM. mRNA expression of key genes involved in ECM turnover was significantly different when comparing primary human intestinal myofibroblasts cultured in 3D scaffolds with those cultured in standard 2D cultures on plastic dishes. Moreover, incubation with key profibrogenic growth factors such as TGFβ1 and PDGF-BB resulted in markedly different effects in standard 2D vs 3D cultures, further emphasizing the importance of using 3D cell cultures. Conclusions These results confirm the feasibility of 3D culture of human intestinal myofibroblasts in intestinal ECM scaffolds as an innovative platform for disease modeling, biomarker discovery, and drug testing in intestinal fibrosis.

scholarly journals Spheroids as a Type of Three-Dimensional Cell Cultures—Examples of Methods of Preparation and the Most Important Application

2020 ◽  
Vol 21 (17) ◽  
pp. 6225 ◽  
Author(s):  
Kamila Białkowska ◽  
Piotr Komorowski ◽  
Maria Bryszewska ◽  
Katarzyna Miłowska
Keyword(s):  
Cell Cultures ◽  
Cell Biology ◽  
Disease Modeling ◽  
Three Dimensional ◽  
3D Culture ◽  
Matrix Interactions ◽  
Vitro Model ◽  
Extracellular Matrix Interactions ◽  
Natural Cell

Cell cultures are very important for testing materials and drugs, and in the examination of cell biology and special cell mechanisms. The most popular models of cell culture are two-dimensional (2D) as monolayers, but this does not mimic the natural cell environment. Cells are mostly deprived of cell–cell and cell–extracellular matrix interactions. A much better in vitro model is three-dimensional (3D) culture. Because many cell lines have the ability to self-assemble, one 3D culturing method is to produce spheroids. There are several systems for culturing cells in spheroids, e.g., hanging drop, scaffolds and hydrogels, and these cultures have their applications in drug and nanoparticles testing, and disease modeling. In this paper we would like to present methods of preparation of spheroids in general and emphasize the most important applications.

scholarly journals Quantitative Image-Based Cell Viability (QuantICV) Assay for Microfluidic 3D Tissue Culture Applications

Micromachines ◽  
2020 ◽  
Vol 11 (7) ◽  
pp. 669
Author(s):  
Louis Jun Ye Ong ◽  
Liang Zhu ◽  
Gabriel Jenn Sern Tan ◽  
Yi-Chin Toh
Keyword(s):  
Tissue Culture ◽  
Cell Viability ◽  
Cell Cultures ◽  
Drug Testing ◽  
3D Culture ◽  
Liquid Volume ◽  
Small Scale ◽  
Tissue Cultures ◽  
Cell Viability Assay ◽  
Quantitative Image

Microfluidic 3D tissue culture systems are attractive for in vitro drug testing applications due to the ability of these platforms to generate 3D tissue models and perform drug testing at a very small scale. However, the minute cell number and liquid volume impose significant technical challenges to perform quantitative cell viability measurements using conventional colorimetric or fluorometric assays, such as MTS or Alamar Blue. Similarly, live-dead staining approaches often utilize metabolic dyes that typically label the cytoplasm of live cells, which makes it difficult to segment and count individual cells in compact 3D tissue cultures. In this paper, we present a quantitative image-based cell viability (QuantICV) assay technique that circumvents current challenges of performing the quantitative cell viability assay in microfluidic 3D tissue cultures. A pair of cell-impermeant nuclear dyes (EthD-1 and DAPI) were used to sequentially label the nuclei of necrotic and total cell populations, respectively. Confocal microscopy and image processing algorithms were employed to visualize and quantify the cell nuclei in the 3D tissue volume. The QuantICV assay was validated and showed good concordance with the conventional bulk MTS assay in static 2D and 3D tumor cell cultures. Finally, the QuantICV assay was employed as an on-chip readout to determine the differential dose responses of parental and metastatic 3D oral squamous cell carcinoma (OSCC) to Gefitinib in a microfluidic 3D culture device. This proposed technique can be useful in microfluidic cell cultures as well as in a situation where conventional cell viability assays are not available.

scholarly journals Proteomics of Colorectal Cancer: Tumors, Organoids, and Cell Cultures—A Minireview

2020 ◽  
Vol 7 ◽  
Author(s):  
Philip H. Lindhorst ◽  
Amanda B. Hummon
Keyword(s):  
Colorectal Cancer ◽  
Cancer Research ◽  
Cell Cultures ◽  
Three Dimensional ◽  
Protein Composition ◽  
Model Systems ◽  
Advantages And Disadvantages ◽  
3D Cell Cultures ◽  
Important Field ◽  
Complete Protein

Proteomics, the study of the complete protein composition of a sample, is an important field for cancer research. Changes in the proteome can serve as a biomarker of cancer or lead to the development of a targeted therapy. This minireview will focus on mass spectrometry-based proteomics studies applied specifically to colorectal cancer, particularly the variety of cancer model systems used, including tumor samples, two-dimensional (2D) and three-dimensional (3D) cell cultures such as spheroids and organoids. A thorough discussion of the application of these systems will accompany the review of the literature, as each provides distinct advantages and disadvantages for colorectal cancer research. Finally, we provide conclusions and future perspectives for the application of these model systems to cancer research as a whole.

The Use of Hydrogels as Biomimetic Materials for 3D Cell Cultures

2017 ◽  
Vol 70 (1) ◽  
pp. 1 ◽  
Author(s):  
Eric Y. Du ◽  
Adam D. Martin ◽  
Celine Heu ◽  
Pall Thordarson
Keyword(s):  
Cell Culture ◽  
Cell Cultures ◽  
Three Dimensional ◽  
3D Culture ◽  
Biomimetic Materials ◽  
Cell Culture Techniques ◽  
Culture Techniques ◽  
3D Cell Cultures ◽  
Recent Developments ◽  
A Cell

With the recent developments in cell cultures and biomimetic materials, there is growing evidence indicating that long-established two-dimensional (2D) cell culture techniques are slowly being phased out and replaced with three-dimensional (3D) cell cultures. This is due to the 3D cell cultures better mimicking the natural extracellular matrix (ECM) where cells are found. The emergence of self-assembled hydrogels as an ECM mimic has revolutionised the field owing to their ability to closely simulate the fibrous nature of the ECM. Here, we review recent progress in using hydrogels as biomimetic materials in 3D cell cultures, particularly supramolecular peptide hydrogels. With greater comprehension of the behaviour of cells in these hydrogels, a cell culture system that can be used in a wide array of 3D culture-based applications can be developed.

scholarly journals Dual-matrix 3D culture system as a biomimetic model of epithelial tissues

2019 ◽  
Author(s):  
Diana Bogorodskaya ◽  
Joshua S. McLane ◽  
Lee A. Ligon
Keyword(s):  
Cell Biology ◽  
Culture System ◽  
Drug Testing ◽  
Disease Modeling ◽  
Wide Spectrum ◽  
3D Culture ◽  
Cell Types ◽  
3D Cultures ◽  
3D Culture System

ABSTRACTRecent years have seen an unprecedented rise in the use of 3D culture systems, both in fundamental research and in more translational settings such as drug testing and disease modeling. However, 3D cultures often remain underused by cell biology labs, both due to technical difficulties in system setup and inherent drawbacks of many of the common systems. Here we describe an easy to use, inexpensive and rapidly assembled 3D culture system, suitable for generation of both normal polarized epithelial cysts and in-situ tumor spheroids. This system allows for exploration of many questions of normal and cancer cell biology, including morphogenesis, epithelial polarization, cell motility, intra- and intercellular communication, invasion, metastasis, and tumor-stoma interaction. The 3D cultures are made up of a stiffness tunable, dual-matrix model that can incorporate co-culture of multiple cell types. The model allows for increased physiological relevance by mimicking the organization, ligand composition and stiffness presentin-vivo. The setup allows for a wide spectrum of manipulation, including removing cells from the system for DNA/protein expression, transfection and high-resolution imaging of live or fixed cells.

scholarly journals Three-Dimensional Cell Cultures in Drug Discovery and Development

2017 ◽  
Vol 22 (5) ◽  
pp. 456-472 ◽  
Author(s):  
Ye Fang ◽  
Richard M. Eglen
Keyword(s):  
Drug Discovery ◽  
Cell Cultures ◽  
Disease Modeling ◽  
Three Dimensional ◽  
3D Cell Culture ◽  
Compound Identification ◽  
3D Cell Cultures ◽  
Dimensional Cell ◽  
Early Drug

The past decades have witnessed significant efforts toward the development of three-dimensional (3D) cell cultures as systems that better mimic in vivo physiology. Today, 3D cell cultures are emerging, not only as a new tool in early drug discovery but also as potential therapeutics to treat disease. In this review, we assess leading 3D cell culture technologies and their impact on drug discovery, including spheroids, organoids, scaffolds, hydrogels, organs-on-chips, and 3D bioprinting. We also discuss the implementation of these technologies in compound identification, screening, and development, ranging from disease modeling to assessment of efficacy and safety profiles.

Application of Ultrasound Elastography for Assessing Intestinal Fibrosis in Inflammatory Bowel Disease: Fiction or Reality?

2020 ◽  
Vol 21 ◽  
Author(s):  
Roberto Gabbiadini ◽  
Eirini Zacharopoulou ◽  
Federica Furfaro ◽  
Vincenzo Craviotto ◽  
Alessandra Zilli ◽  
...  
Keyword(s):  
Inflammatory Bowel Disease ◽  
Sample Size ◽  
Bowel Disease ◽  
Small Sample Size ◽  
Shear Wave Elastography ◽  
Small Sample ◽  
Ultrasound Elastography ◽  
Invasive Technique ◽  
Intestinal Fibrosis ◽  
Inflammatory Bowel

Background: Intestinal fibrosis and subsequent strictures represent an important burden in inflammatory bowel disease (IBD). The detection and evaluation of the degree of fibrosis in stricturing Crohn’s disease (CD) is important to address the best therapeutic strategy (medical anti-inflammatory therapy, endoscopic dilation, surgery). Ultrasound elastography (USE) is a non-invasive technique that has been proposed in the field of IBD for evaluating intestinal stiffness as a biomarker of intestinal fibrosis. Objective: The aim of this review is to discuss the ability and current role of ultrasound elastography in the assessment of intestinal fibrosis. Results and Conclusion: Data on USE in IBD are provided by pilot and proof-of-concept studies with small sample size. The first type of USE investigated was strain elastography, while shear wave elastography has been introduced lately. Despite the heterogeneity of the methods of the studies, USE has been proven to be able to assess intestinal fibrosis in patients with stricturing CD. However, before introducing this technique in current practice, further studies with larger sample size and homogeneous parameters, testing reproducibility, and identification of validated cut-off values are needed.

scholarly journals Can keratin scaffolds be used for creating three-dimensional cell cultures?

Open Medicine ◽  
2020 ◽  
Vol 15 (1) ◽  
pp. 249-253
Author(s):  
Marta Bochynska-Czyz ◽  
Patrycja Redkiewicz ◽  
Hanna Kozlowska ◽  
Joanna Matalinska ◽  
Marek Konop ◽  
...  
Keyword(s):  
Cell Cultures ◽  
Chemical Activation ◽  
Three Dimensional ◽  
Enzymatic Digestion ◽  
Pepsin Digestion ◽  
Cell Culturing ◽  
3D Cell Cultures ◽  
Associated Proteins ◽  
Positive Effect ◽  
Dimensional Cell

AbstractThree-dimensional (3D) cell cultures were created with the use of fur keratin associated proteins (F-KAPs) as scaffolds. The procedure of preparation F-KAP involves combinations of chemical activation and enzymatic digestion. The best result in porosity and heterogeneity of F-KAP surface was received during pepsin digestion. The F-KAP had a stable structure, no changes were observed after heat treatment, shaking and washing. The 0.15-0.5 mm fraction had positive effect for formation of 3D scaffolds and cell culturing. Living rat mesenchymal cells on the F-KAP with no abnormal morphology were observed by SEM during 32 days of cell culturing.

scholarly journals Correction: Assessment of the toxicity and inflammatory effects of different-sized zinc oxide nanoparticles in 2D and 3D cell cultures

RSC Advances ◽  
2020 ◽  
Vol 10 (72) ◽  
pp. 44397-44397
Author(s):  
Zhipan Wu ◽  
Rongfa Guan ◽  
Miao Tao ◽  
Fei Lyu ◽  
Guozhou Cao ◽  
...  
Keyword(s):  
Zinc Oxide ◽  
Cell Cultures ◽  
Zinc Oxide Nanoparticles ◽  
Oxide Nanoparticles ◽  
3D Cell Cultures ◽  
2D And 3D

Correction for ‘Assessment of the toxicity and inflammatory effects of different-sized zinc oxide nanoparticles in 2D and 3D cell cultures’ by Zhipan Wu, Rongfa Guan, Miao Tao et al., RSC Adv., 2017, 7, 12437–12445, DOI: 10.1039/C6RA27334C.

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.

Sign in / Sign up

Export Citation Format

Share Document