Novel pan sigma receptor modulator exerts strong apoptotic effect in in vitro primary 3D-cell cultures of human glioblastoma

These unprecedented times have forced the scientific community to gather to face the COVID-19 pandemic. Efforts in diverse directions have been made. A multi-university team has focused on the identification of the host (human) proteins interacting with SARS-CoV-2 viral proteins, with the aim of hampering these interactions that may cause severe COVID-19 symptoms. Sigma-1 and sigma-2 receptors surprisingly belong to the “druggable” host proteins found, with the pan-sigma receptor modulator PB28 displaying the most potent anti–SARS-CoV-2 activity in in vitro assays. Being 20-fold more active than hydroxychloroquine, without cardiac side effects, PB28 is a promising antiviral candidate worthy of further investigation. Our research group developed PB28 in 1996 and have thoroughly characterized its biological properties since then. Structure–affinity relationship (SAfiR) studies at the sigma receptor subtypes were also undertaken with PB28 as the lead compound. We herein report our knowledge of PB28 to share information that may help to gain insight into the antiviral action of this compound and sigma receptors, while providing structural hints that may speed up the translation into therapeutics of this class of ligands.

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Ratiometric Nanoviscometers: Applications for Measuring Cellular Physical Properties in 3D Cultures

SLAS TECHNOLOGY Translating Life Sciences Innovation ◽

10.1177/2472630319901262 ◽

2020 ◽

Vol 25 (3) ◽

pp. 234-246

Author(s):

Charles McRae White ◽

Mark A. Haidekker ◽

William S. Kisaalita

Keyword(s):

Cell Culture ◽

Cell Cultures ◽

Cell Biology ◽

Low Cost ◽

3D Cell Culture ◽

Biomechanical Properties ◽

Practical Applications ◽

3D Cell Cultures

New insights into the biomechanical properties of cells are revealing the importance of these properties and how they relate to underlying molecular, architectural, and behavioral changes associated with cell state and disease processes. However, the current understanding of how these in vitro biomechanical properties are associated with in vivo processes has been developed based on the traditional monolayer (two-dimensional [2D]) cell culture, which traditionally has not translated well to the three-dimensional (3D) cell culture and in vivo function. Many gold standard methods and tools used to observe the biomechanical properties of 2D cell cultures cannot be used with 3D cell cultures. Fluorescent molecules can respond to external factors almost instantaneously and require relatively low-cost instrumentation. In this review, we provide the background on fluorescent molecular rotors, which are attractive tools due to the relationship of their emission quantum yield with environmental microviscosity. We make the case for their use in both 2D and 3D cell cultures and speculate on their fundamental and practical applications in cell biology.

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Batch Fabrication of Three-Dimensional Microelectrode Arrays For The In-Vitro Investigation Of Tissue Slices And 3D Cell Cultures

Frontiers in Neuroscience ◽

10.3389/conf.fnins.2016.93.00133 ◽

2016 ◽

Vol 10 ◽

Author(s):

Martina Manuel ◽

Klaus Stefan ◽

Stett Alfred ◽

Herrmann Thoralf ◽

Zeck G�nther ◽

...

Keyword(s):

Cell Cultures ◽

Three Dimensional ◽

Microelectrode Arrays ◽

Tissue Slices ◽

Batch Fabrication ◽

3D Cell Cultures ◽

In Vitro Investigation

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Precise, high-throughput production of multicellular spheroids with a bespoke 3D bioprinter

10.1101/2020.04.06.028548 ◽

2020 ◽

Cited By ~ 1

Author(s):

Robert H. Utama ◽

Lakmali Atapattu ◽

Aidan P. O’Mahony ◽

Christopher M. Fife ◽

Jongho Baek ◽

...

Keyword(s):

Cell Cultures ◽

High Throughput ◽

Cell Number ◽

Drug Dose ◽

3D Bioprinting ◽

Multicellular Spheroids ◽

Drop On Demand ◽

3D Cell Cultures ◽

The Impact

Abstract3D in vitro cancer models are important therapeutic and biological discovery tools, yet formation of multicellular spheroids in a throughput and highly controlled manner to achieve robust and statistically relevant data, remains challenging. Here, we developed an enabling technology consisting of a bespoke drop-on-demand 3D bioprinter capable of high-throughput printing of 96-well plates of spheroids. 3D-multicellular spheroids are embedded inside a tissue-like matrix with precise control over size and cell number. Application of 3D bioprinting for high-throughput drug screening was demonstrated with doxorubicin. Measurements showed that IC50 values were sensitive to spheroid size, embedding and how spheroids conform to the embedding, revealing parameters shaping biological responses in these models. Our study demonstrates the potential of 3D bioprinting as a robust high-throughput platform to screen biological and therapeutic parameters.Significance StatementIn vitro 3D cell cultures serve as more realistic models, compared to 2D cell culture, for understanding diverse biology and for drug discovery. Preparing 3D cell cultures with defined parameters is challenging, with significant failure rates when embedding 3D multicellular spheroids into extracellular mimics. Here, we report a new 3D bioprinter we developed in conjunction with bioinks to allow 3D-multicellular spheroids to be produced in a high-throughput manner. High-throughput production of embedded multicellular spheroids allowed entire drug-dose responses to be performed in 96-well plate format with statistically relevant numbers of data points. We have deconvoluted important parameters in drug responses including the impact of spheroid size and embedding in an extracellular matrix mimic on IC50 values.

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An Interpenetrating Alginate/Gelatin Network for Three-Dimensional (3D) Cell Cultures and Organ Bioprinting

Molecules ◽

10.3390/molecules25030756 ◽

2020 ◽

Vol 25 (3) ◽

pp. 756 ◽

Cited By ~ 4

Author(s):

Qiuhong Chen ◽

Xiaohong Tian ◽

Jun Fan ◽

Hao Tong ◽

Qiang Ao ◽

...

Keyword(s):

Cell Cultures ◽

Structural Integrity ◽

Polymer Network ◽

Three Dimensional ◽

Biochemical Properties ◽

Interpenetrating Polymer Network ◽

3D Cell Cultures ◽

Wide Range ◽

The Individual

Crosslinking is an effective way to improve the physiochemical and biochemical properties of hydrogels. In this study, we describe an interpenetrating polymer network (IPN) of alginate/gelatin hydrogels (i.e., A-G-IPN) in which cells can be encapsulated for in vitro three-dimensional (3D) cultures and organ bioprinting. A double crosslinking model, i.e., using Ca2+ to crosslink alginate molecules and transglutaminase (TG) to crosslink gelatin molecules, is exploited to improve the physiochemical, such as water holding capacity, hardness and structural integrity, and biochemical properties, such as cytocompatibility, of the alginate/gelatin hydrogels. For the sake of convenience, the individual ionic (i.e., only treatment with Ca2+) or enzymatic (i.e., only treatment with TG) crosslinked alginate/gelatin hydrogels are referred as alginate-semi-IPN (i.e., A-semi-IPN) or gelatin-semi-IPN (i.e., G-semi-IPN), respectively. Tunable physiochemical and biochemical properties of the hydrogels have been obtained by changing the crosslinking sequences and polymer concentrations. Cytocompatibilities of the obtained hydrogels are evaluated through in vitro 3D cell cultures and bioprinting. The double crosslinked A-G-IPN hydrogel is a promising candidate for a wide range of biomedical applications, including bioartificial organ manufacturing, high-throughput drug screening, and pathological mechanism analyses.

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Topical protection of human esophageal mucosal integrity

AJP Gastrointestinal and Liver Physiology ◽

10.1152/ajpgi.00424.2014 ◽

2015 ◽

Vol 308 (12) ◽

pp. G975-G980 ◽

Cited By ~ 20

Author(s):

P. Woodland ◽

F. Batista-Lima ◽

C. Lee ◽

S. L. Preston ◽

P. Dettmar ◽

...

Keyword(s):

Cell Cultures ◽

Reflux Disease ◽

Esophageal Mucosa ◽

Apical Surface ◽

Alginate Solution ◽

Ussing Chambers ◽

Mucosal Integrity ◽

3D Cell Cultures ◽

Induced Changes

Patients with nonerosive reflux disease exhibit impaired esophageal mucosal integrity, which may underlie enhanced reflux perception. In vitro topical application of an alginate solution can protect mucosal biopsies against acid-induced changes in transepithelial electrical resistance (TER). We aimed to confirm this finding in a second model using 3D cell cultures and to assess prolonged protection in a biopsy model. We assessed the protective effect of a topically applied alginate solution 1 h after application. 3D cell cultures were grown by using an air-liquid interface and were studied in Ussing chambers. The apical surface was “protected” with 200 μl of either alginate or viscous control or was unprotected. The tissue was exposed to pH 3 + bile acid solution for 30 min and TER change was calculated. Distal esophageal mucosal biopsies were taken from 12 patients and studied in Ussing chambers. The biopsies were coated with either alginate or viscous control solution. The biopsies were then bathed in pH 7.4 solution for 1 h. The luminal chamber solution was replaced with pH 2 solution for 30 min. Percentage changes in TER were recorded. In five biopsies fluorescein-labeled alginate solution was used to allow immunohistological localization of the alginate after 1 h. In the cell culture model, alginate solution protected tissue against acid-induced change in TER. In biopsies, 60 min after protection with alginate solution, the acidic exposure caused a −8.3 ± 2.2% change in TER compared with −25.1 ± 4.5% change after protection with the viscous control ( P < 0.05). Labeled alginate could be seen coating the luminal surface in all cases. In vitro, alginate solutions can adhere to the esophageal mucosa for up to 1 h and exert a topical protectant effect. Durable topical protectants can be further explored as first-line/add-on therapies for gastroesophageal reflux disease.

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