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three-dimensional culture

Ghasem Barati, Samad Nadri, Ramin Hajian, Ali Rahmani, Hossein Mostafavi, Yousef Mortazavi, Amir Hossein Taromchi
Tissue and stem cell encapsulation andtransplantation were considered as promising tools in the treatment of patients with diabetes mellitus. The aim of this study was to evaluate the effect of microfluidic encapsulation on the differentiation of trabecular meshwork mesenchymal stem cells (TM-MSC), into insulin-producing cells (IPCs) both in vitro and in vivo. The presence of differentiated cells in microfibers (three dimensional [3D]) and tissue culture plates (TCPS; two dimensional [2D]) culture was evaluated by detecting mRNA and protein expression of pancreatic islet-specific markers as well as measuring insulin release of cells in response to glucose challenges...
October 14, 2018: Journal of Cellular Physiology
Mark C Allenby, Nicki Panoskaltsis, Asma Tahlawi, Susana Brito Dos Santos, Athanasios Mantalaris
Traditional culture systems for human erythropoiesis lack microenvironmental niches, spatial marrow gradients and dense cellularity rendering them incapable of effectively translating marrow physiology ex vivo. Herein, a bio-inspired three-dimensional (3D) perfusion bioreactor was engineered and inoculated with unselected single donor umbilical cord blood mononuclear cells (CBMNCs). Functional stromal and hematopoietic environments supporting long-term erythropoiesis were generated using defined medium supplemented only with stem cell factor (SCF) and erythropoietin (EPO) at near physiological concentrations...
August 8, 2018: Biomaterials
Kristin Robin Ko, Meng-Chiao Tsai, John P Frampton
Extracellular matrix-based hydrogels such as Matrigel are easy-to-use, commercially available, and offer environments for three-dimensional (3D) cell culture that mimic native tissue. However, manipulating small volumes of these materials to produce thin-layer 3D culture systems suitable for analysis is difficult due to air-liquid-substrate interfacial tension effects and evaporation. Here, we demonstrate two simple techniques that use standard liquid-handling tools and non-treated 96-well plates to produce uniform, thin-layer constructs for 3D culture of cells in Matrigel...
October 13, 2018: Biotechnology Progress
Marit van Gorsel, Ilaria Elia, Sarah-Maria Fendt
Metabolomics and 13 C tracer analysis are state-of-the-art techniques that allow determining the concentration of metabolites and the activity of metabolic pathways, respectively. Three dimensional (3D) cultures of cancer cells constitute an enriched in vitro environment that can be used to assay anchorage-independent growth, spheroid formation, and extracellular matrix production by (cancer) cells. Here, we describe how to perform metabolomics and 13 C tracer analysis in 3D cultures of cancer cells.
2019: Methods in Molecular Biology
Christian Humpel
Ex vivo cell culture models are of particular interest for neurobiologists, as these allow the study of brain cells in an isolated environment. Organotypic brain slice cultures allow growth of three-dimensional brain tissue that conserves the cellular architecture. This unit describes the preparation and culturing of organotypic brain slices from mice. In three basic protocols, the general procedure, the specific preparation of chopper slices, and slicing of whole-brain vibratome sections are described. Support protocols explain the use of postnatal or adult mice, the preparation of coronal or sagittal slices, the preparation of co-cultures, post-processing of slices, the use of extra filter membranes, and the re-use of membrane inserts...
October 12, 2018: Current Protocols in Immunology
A Martin-Pena, R M Porter, G Plumton, T M McCarrel, A J Morton, M V Guijarro, S C Ghivizzani, B Sharma, G D Palmer
Successful clinical translation of mesenchymal stem cell (MSC)-based therapies for cartilage repair will likely require the implementation of standardised protocols and broadly applicable tools to facilitate the comparisons among cell types and chondroinduction methods. The present study investigated the utility of recombinant lentiviral reporter vectors as reliable tools for comparing chondrogenic potential among primary cell populations and distinguishing cellular-level variations of chondrogenic activity in widely used three-dimensional (3D) culture systems...
October 12, 2018: European Cells & Materials
Elahe Hadavi, Jeroen C H Leijten, Jenny Brinkmann, Pascal Jonkheijm, Marcel Karperien, Aart Alexander van Apeldoorn
Extracellular matrix molecules play significant roles in regulating β-cell function and viability within pancreatic islets by providing mechanical and biological support, stimulating cell survival, proliferation and their endocrine function. During clinical islet transplantation the beta cell's extracellular matrix environment is degraded by enzymatic digestion. Literature suggests that interactions between islet cells and ECM molecules, such as fibronectin, collagen IV and laminin, are essential for maintaining, or stimulation of islet function and survival, and can effect differentiation and proliferation of the endocrine cells...
October 11, 2018: Tissue Engineering. Part C, Methods
Andrea Cerea, Valeria Caprettini, Giulia Bruno, Laura Lovato, Giovanni Melle, Francesco Tantussi, Rosario Capozza, Fabio Moia, Michele Dipalo, Francesco De Angelis
Biological studies on in vitro cell cultures are of fundamental importance to investigate cell response to external stimuli, such as new drugs for the treatment of specific pathologies, or to study communication between electrogenic cells. Although three-dimensional (3D) nanostructures brought tremendous improvements on biosensors used for various biological in vitro studies, including drug delivery and electrical recording, there is still a lack of multifunctional capabilities that could help gain deeper insights in several bio-related research fields...
October 11, 2018: Lab on a Chip
Junko Johzuka, Toshihiro Ona, Masatoshi Nomura
In anti-cancer drug (candidate) screening, there is the need for evaluation at physiological concentrations similar to in vivo. This is often performed by three-dimensionally (3D) cultured cells; however, it requires a long culture period of 2 - 4 weeks with tedious experimental procedures. Here, we report on a high precision surface plasmon resonance (HP-SPR)-3D system. We developed the system with average fluctuation of 50 ndeg s-1 using two-dimensionally cultured cells attached onto a sensor chip by applying collagen on the top to change their activity into in vivo-like conditions without cell division...
2018: Analytical Sciences: the International Journal of the Japan Society for Analytical Chemistry
John A Reid, Peter M Mollica, Robert D Bruno, Patrick C Sachs
BACKGROUND: Standard three-dimensional (3D) in vitro culture techniques, such as those used for mammary epithelial cells, rely on random distribution of cells within hydrogels. Although these systems offer advantages over traditional 2D models, limitations persist owing to the lack of control over cellular placement within the hydrogel. This results in experimental inconsistencies and random organoid morphology. Robust, high-throughput experimentation requires greater standardization of 3D epithelial culture techniques...
October 10, 2018: Breast Cancer Research: BCR
Seungwoo Shin, Jihye Kim, Je-Ryung Lee, Eun-Chae Jeon, Tae-Jin Je, Wonhee Lee, YongKeun Park
Optical diffraction tomography (ODT) enables the reconstruction of the three-dimensional (3D) refractive-index (RI) distribution of a biological cell, which provides invaluable information for cellular and subcellular structures in a non-invasive manner. However, ODT suffers from an inferior axial resolution, due to the limited accessible angles imposed by the numerical aperture of the objective lens. In this study, we propose and experimentally demonstrate an approach to enhance the 3D reconstruction performance in ODT...
October 10, 2018: Lab on a Chip
Guoyi Wu, Shanshan Zhan, Chen Rui, Eiketsu Sho, Xiaolei Shi, Yitao Ding
Hepatocellular carcinoma (HCC) treatments are evaluated by two-dimensional (2D) in vitro culture systems, despite their limited ability to predict drug efficacy. The three-dimensional (3D) microporous scaffold provides the possibility of generating more reliable preclinical models to increase the efficacy of cancer treatments. The physical properties of a microporous cellulosic scaffold were evaluated. The cellulosic scaffold was biocompatible and had a highly porous network with appropriate pore size, swelling rate, and stiffness of cancer cell cultures...
October 9, 2018: Journal of Cellular Biochemistry
Siming Lu, Fabio Cuzzucoli, Jing Jiang, Li-Guo Liang, Yimin Wang, Mengqi Kong, Xin Zhao, Wenguo Cui, Jun Li, ShuQi Wang
Cancer poses a great health threat to both developed and developing countries, and anti-cancer drugs are of important interest for improved clinical outcomes. Although tumor-on-a-chip technologies offer a feasible approach to screening drug toxicity, their capability to mimic the native tumor microenvironment (TME) is still limited. For better mimicry of the TME, we developed a biomimetic three-dimensional (3D) liver tumor-on-a-chip with the integration of essential components derived from decellularized liver matrix (DLM) with gelatin methacryloyl (GelMA) in a microfluidics-based 3D dynamic cell culture system...
October 9, 2018: Lab on a Chip
Francisco Javier Ortiz-López, Elsa Alcalde, Aida Sarmiento-Vizcaíno, Caridad Díaz, Bastien Cautain, Luis A García, Gloria Blanco, Fernando Reyes
Fractionation of the bioactive extract of a culture of the marine derived actinomycete Streptomyces cyaneofuscatus M-157 led to the isolation of the known 3-hydroxyquinaldic acid ( 4 ), its amide ( 5 ) and three new derivatives ( 1 ⁻ 3 ) containing different amino acid residues. The structures of the new molecules ( 1 ⁻ 3 ), including their absolute configuration, were determined by the analysis of their ESI-TOF MS and one-dimensional (1D) and two-dimensional (2D) NMR spectra and advanced Marfey's analysis of their hydrolyzation products...
October 8, 2018: Marine Drugs
I González-Domínguez, L Cervera, F Gòdia, M Roldán
Although cationic polymers are widely used for DNA delivery, the relationship between the properties of the formed complexes and their biological activity is not fully understood. Here, we propose a novel procedure consisting of superresolved images coupled with quantitative colocalization to analyse DNA release in living cells. This work compares the different workflows available in a quantitative colocalization study of DNA delivery using polyethylenimine as transfection reagent. A nimble workflow with deconvolution in three-dimensional images was developed...
October 8, 2018: Journal of Microscopy
Toshihiro Sera, Shingo Komine, Masataka Arai, Yasuhiro Sunaga, Hideo Yokota, Susumu Kudo
Intracellular and intercellular Ca2+ waves play key roles in cellular functions, and focal stimulation triggers Ca2+ wave propagation from stimulation points to neighboring cells, involving localized metabolism reactions and specific diffusion processes. Among these, inositol 1,4,5-trisphosphate (IP3 ) is produced at membranes and diffuses into the cytoplasm to release Ca2+ from endoplasmic reticulum (ER). In this study, we developed a three-dimensional (3D) simulation model for intercellular and intracellular Ca2+ waves in endothelial cells (ECs)...
October 5, 2018: Biochemical and Biophysical Research Communications
Yan Xiao, Miaomiao Zhou, Mi Zhang, Wei Liu, Yan Zhou, Meidong Lang
The development of three-dimensional (3D) porous scaffolds for soft tissue engineering mainly focused on manipulation of scaffold properties with cell behaviors. By emulsion freeze-drying method, four types of porous scaffolds were prepared from amorphous poly(4-methy-ε-caprolactone) (PMCL) and semi-crystalline poly(ε-caprolactone) (PCL) at different weight ratios, named as PMCL0, PMCL30, PMCL50 and PMCL70, respectively. Visual observation on cross-sectional images of the scaffolds appeared as sponge-like materials with three-dimensional and highly porous morphologies...
September 28, 2018: Colloids and Surfaces. B, Biointerfaces
Zhuosi Li, Osamu Kurosawa, Hiroo Iwata
We developed a novel engineering technique to induce differentiation of human induced pluripotent stem cells (hiPSCs) into organoids mimicking the trophectoderm (TE). Here, hiPSCs were cultured on a limited area of 2-4 mm in diameter. After 15-20 days, spherical cysts appeared on the surface of the limited area. Secretion of human chorionic gonadotrophin (hCG) began to increase after ∼ 20 days and remained dramatically elevated over the next 20 days. Limited-area-cultured cysts exhibited expression of hCG, which was a result of epithelial differentiation...
October 3, 2018: Biochemical and Biophysical Research Communications
Shinkichi Ishikawa, Kazushi Matsumura, Nobumasa Kitamura, Yuichiro Takanami, Shigeaki Ito
Cigarette smoke (CS) is a major risk factor in the development of chronic inflammatory lung diseases such as chronic obstructive pulmonary disease. A comprehensive investigation of the biological impacts of chronic CS exposure on lung tissue is therefore important for understanding the pathogenesis of lung disease. We used three-dimensional (3D) organotypic human bronchial tissue cultures and metabolomics, transcriptomics, and proteomics to investigate changes in biological processes affected by repeated whole-CS exposure...
October 3, 2018: Toxicology in Vitro: An International Journal Published in Association with BIBRA
Keykavos Gholami, Gholamreza Pourmand, Morteza Koruji, Mohammadali Sadighigilani, Shadan Navid, Fariborz Izadyar, Mehdi Abbasi
Optimization of in vitro culture system for the expansion and the maturation of male germ cells to post meiotic stages is a valuable tool for studies exploring spermatogenesis regulation and the management of male infertility. Several studies have reported promising results of mouse spermatogonial stem cells culture in three-dimensional (3D) culture systems and a subsequent production of sperm. In the present study, we investigated the capacity of a three-dimensional soft agar culture system (SACS) supplemented with Knockout Serum Replacement (KSR) in colony formation and inducing human germ cells to reach post-meiotic stages...
October 2, 2018: Reproductive Biology
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