scholarly journals Three-Dimensional Biofabrication Models of Endometriosis and the Endometriotic Microenvironment

2020 ◽  
Author(s):  
Jillian R. H. Wendel ◽  
Xiyin Wang ◽  
Lester J. Smith ◽  
Shannon M. Hawkins
Keyword(s):  
Cell Line ◽  
Three Dimensional ◽  
Uterine Cavity ◽  
Building Blocks ◽  
Cell Types ◽  
In Vitro Models ◽  
Inflammatory Factors ◽  
Biologically Relevant ◽  
Increased Risk

Endometriosis occurs when endometrial-like tissue grows outside the uterine cavity, leading to pelvic pain, infertility, and increased risk of ovarian cancer. The present study describes the optimization and characterization of cellular spheroids as building blocks for Kenzan scaffold-free method biofabrication and proof-of-concept models of endometriosis and the endometriotic microenvironment. The spheroid building blocks must be a specific diameter (~500 m), compact, round, and smooth to withstand Kenzan biofabrication. Under optimized spheroid conditions for biofabrication, the endometriotic epithelial-like cell line, 12Z, expressed high levels of estrogen-related genes and secreted high amounts of endometriotic inflammatory factors that were independent of TNF stimulation. Heterotypic spheroids, composed of 12Z and T-HESC, an immortalized endometrial stromal cell line, self-assembled into a biologically relevant pattern, consisting of epithelial cells on the outside of the spheroids and stromal cells in the core. 12Z spheroids were biofabricated into large three-dimensional constructs alone, with HEYA8 spheroids, or as heterotypic spheroids with T-HESC. These three-dimensional biofabricated constructs containing multiple monotypic or heterotypic spheroids represent the first scaffold-free biofabricated in vitro models of endometriosis and the endometriotic microenvironment. These efficient and innovative models will allow us to study the complex interactions of multiple cell types within a biologically relevant microenvironment.

scholarly journals Three-Dimensional Biofabrication Models of Endometriosis and the Endometriotic Microenvironment

Biomedicines ◽  
2020 ◽  
Vol 8 (11) ◽  
pp. 525
Author(s):  
Jillian R. H. Wendel ◽  
Xiyin Wang ◽  
Lester J. Smith ◽  
Shannon M. Hawkins
Keyword(s):  
Cell Line ◽  
Three Dimensional ◽  
Uterine Cavity ◽  
Building Blocks ◽  
Cell Types ◽  
In Vitro Models ◽  
Inflammatory Factors ◽  
Biologically Relevant ◽  
Increased Risk

Endometriosis occurs when endometrial-like tissue grows outside the uterine cavity, leading to pelvic pain, infertility, and increased risk of ovarian cancer. The present study describes the optimization and characterization of cellular spheroids as building blocks for Kenzan scaffold-free method biofabrication and proof-of-concept models of endometriosis and the endometriotic microenvironment. The spheroid building blocks must be of a specific diameter (~500 μm), compact, round, and smooth to withstand Kenzan biofabrication. Under optimized spheroid conditions for biofabrication, the endometriotic epithelial-like cell line, 12Z, expressed high levels of estrogen-related genes and secreted high amounts of endometriotic inflammatory factors that were independent of TNFα stimulation. Heterotypic spheroids, composed of 12Z and T-HESC, an immortalized endometrial stromal cell line, self-assembled into a biologically relevant pattern, consisting of epithelial cells on the outside of the spheroids and stromal cells in the core. 12Z spheroids were biofabricated into large three-dimensional constructs alone, with HEYA8 spheroids, or as heterotypic spheroids with T-HESC. These three-dimensional biofabricated constructs containing multiple monotypic or heterotypic spheroids represent the first scaffold-free biofabricated in vitro models of endometriosis and the endometriotic microenvironment. These efficient and innovative models will allow us to study the complex interactions of multiple cell types within a biologically relevant microenvironment.

scholarly journals Three-dimensional testicular organoids as novel in vitro models of testicular biology and toxicology

2019 ◽  
Vol 5 (3) ◽  
Author(s):  
Sadman Sakib ◽  
Anna Voigt ◽  
Taylor Goldsmith ◽  
Ina Dobrinski
Keyword(s):  
Reproductive Biology ◽  
Monolayer Culture ◽  
Three Dimensional ◽  
Cell Types ◽  
In Vitro Models ◽  
Toxicity Screening ◽  
Current State ◽  
Potential Applications ◽  
Multiple Cell

Abstract Organoids are three dimensional structures consisting of multiple cell types that recapitulate the cellular architecture and functionality of native organs. Over the last decade, the advent of organoid research has opened up many avenues for basic and translational studies. Following suit of other disciplines, research groups working in the field of male reproductive biology have started establishing and characterizing testicular organoids. The three-dimensional architectural and functional similarities of organoids to their tissue of origin facilitate study of complex cell interactions, tissue development and establishment of representative, scalable models for drug and toxicity screening. In this review, we discuss the current state of testicular organoid research, their advantages over conventional monolayer culture and their potential applications in the field of reproductive biology and toxicology.

scholarly journals In vitro models of the human heart

Development ◽  
2021 ◽  
Vol 148 (16) ◽  
Author(s):  
Pablo Hofbauer ◽  
Stefan M. Jahnel ◽  
Sasha Mendjan
Keyword(s):  
Heart Development ◽  
Human Heart ◽  
Three Dimensional ◽  
Cell Types ◽  
In Vitro Models ◽  
Major Bottleneck ◽  
Key Aspects ◽  
New Generation ◽  
Self Organizing

ABSTRACT Cardiac congenital disabilities are the most common organ malformations, but we still do not understand how they arise in the human embryo. Moreover, although cardiovascular disease is the most common cause of death globally, the development of new therapies is lagging compared with other fields. One major bottleneck hindering progress is the lack of self-organizing human cardiac models that recapitulate key aspects of human heart development, physiology and disease. Current in vitro cardiac three-dimensional systems are either engineered constructs or spherical aggregates of cardiomyocytes and other cell types. Although tissue engineering enables the modeling of some electro-mechanical properties, it falls short of mimicking heart development, morphogenetic defects and many clinically relevant aspects of cardiomyopathies. Here, we review different approaches and recent efforts to overcome these challenges in the field using a new generation of self-organizing embryonic and cardiac organoids.

scholarly journals Development of In Vitro Corneal Models: Opportunity for Pharmacological Testing

2020 ◽  
Vol 3 (4) ◽  
pp. 74
Author(s):  
Valentina Citi ◽  
Eugenia Piragine ◽  
Simone Brogi ◽  
Sara Ottino ◽  
Vincenzo Calderone
Keyword(s):  
Three Dimensional ◽  
Toxicity Testing ◽  
Cell Types ◽  
In Vitro Models ◽  
Rapid Expansion ◽  
Anatomy And Physiology ◽  
Complex Anatomy ◽  
Computational Procedures ◽  
Scientific Challenge

The human eye is a specialized organ with a complex anatomy and physiology, because it is characterized by different cell types with specific physiological functions. Given the complexity of the eye, ocular tissues are finely organized and orchestrated. In the last few years, many in vitro models have been developed in order to meet the 3Rs principle (Replacement, Reduction and Refinement) for eye toxicity testing. This procedure is highly necessary to ensure that the risks associated with ophthalmic products meet appropriate safety criteria. In vitro preclinical testing is now a well-established practice of significant importance for evaluating the efficacy and safety of cosmetic, pharmaceutical, and nutraceutical products. Along with in vitro testing, also computational procedures, herein described, for evaluating the pharmacological profile of potential ocular drug candidates including their toxicity, are in rapid expansion. In this review, the ocular cell types and functionality are described, providing an overview about the scientific challenge for the development of three-dimensional (3D) in vitro models.

scholarly journals Cerebral Pericytes and Endothelial Cells Communicate through Inflammasome-Dependent Signals

2021 ◽  
Vol 22 (11) ◽  
pp. 6122
Author(s):  
Mihály Kozma ◽  
Ádám Mészáros ◽  
Ádám Nyúl-Tóth ◽  
Kinga Molnár ◽  
Laura Costea ◽  
...  
Keyword(s):  
Endothelial Cells ◽  
Neurovascular Unit ◽  
Barrier Properties ◽  
Cell Types ◽  
In Vitro Models ◽  
Canonical Pathway ◽  
Inflammatory Factors ◽  
Pyrin Domain ◽  
Inflammatory Stimuli

By upregulation of cell adhesion molecules and secretion of proinflammatory cytokines, cells of the neurovascular unit, including pericytes and endothelial cells, actively participate in neuroinflammatory reactions. As previously shown, both cell types can activate inflammasomes, cerebral endothelial cells (CECs) through the canonical pathway, while pericytes only through the noncanonical pathway. Using complex in vitro models, we demonstrate here that the noncanonical inflammasome pathway can be induced in CECs as well, leading to a further increase in the secretion of active interleukin-1β over that observed in response to activation of the canonical pathway. In parallel, a more pronounced disruption of tight junctions takes place. We also show that CECs respond to inflammatory stimuli coming from both the apical/blood and the basolateral/brain directions. As a result, CECs can detect factors secreted by pericytes in which the noncanonical inflammasome pathway is activated and respond with inflammatory activation and impairment of the barrier properties. In addition, upon sensing inflammatory signals, CECs release inflammatory factors toward both the blood and the brain sides. Consequently, CECs activate pericytes by upregulating their expression of NLRP3 (NOD-, LRR-, and pyrin domain-containing protein 3), an inflammasome-forming pattern recognition receptor. In conclusion, cerebral pericytes and endothelial cells mutually activate each other in inflammation.

A Human Corneal Equivalent Constructed from SV40-immortalised Corneal Cell Lines

2005 ◽  
Vol 33 (1) ◽  
pp. 37-45 ◽  
Author(s):  
Michaela Zorn-Kruppa ◽  
Svitlana Tykhonova ◽  
Gazanfer Belge ◽  
Jürgen Bednarz ◽  
Horst A. Diehl ◽  
...  
Keyword(s):  
Cell Lines ◽  
Cross Sections ◽  
Three Dimensional ◽  
Morphological Characteristics ◽  
Drug Efficacy ◽  
Collagen Matrix ◽  
Cell Types ◽  
In Vitro Models ◽  
Equivalent Model

Within the last decade, extensive research in the field of tissue and organ engineering has focused on the development of in vitro models of the cornea. The use of organotypic, three-dimensional corneal equivalents has several advantages over simple monolayer cultures. The aim of this study was to develop a corneal equivalent model composed of the same cell types as in the natural human tissue, but by using immortalised cell lines to ensure reproducibility and to minimise product variation. We report our success in the establishment of an SV40-immortalised human corneal keratocyte cell line (designated HCK). A collagen matrix, built up with these cells, displayed the morphological characteristics of the human stromal tissue and served as a biomatrix for the immortalised human corneal epithelial and endothelial cells. Histological cross-sections of the whole-cornea equivalents resemble human corneas in tissue structure. This organotypic in vitro model may serve as a research tool for the ophthalmic science community, as well as a model system for testing for eye irritancy and drug efficacy.

scholarly journals Development of In Vitro Corneal Models: Opportunity for Pharmacological Testing and Legislative Aspects

2020 ◽  
Author(s):  
Valentina Citi ◽  
Eugenia Piragine ◽  
Simone Brogi ◽  
Sara Ottino ◽  
Marco Sansò ◽  
...  
Keyword(s):  
Three Dimensional ◽  
Toxicity Testing ◽  
Cell Types ◽  
In Vitro Models ◽  
Rapid Expansion ◽  
Anatomy And Physiology ◽  
Complex Anatomy ◽  
Computational Procedures ◽  
Scientific Challenge

Human eye is a specialized organ with complex anatomy and physiology, because it is characterized by different cell types with specific physiological functions. Given the complexity of the eye, ocular tissues are finely organized and orchestrated. In the last few years many in vitro models have been developed, in order to meet the 3Rs principle (Replacement, Reduction and Refinement) for eye toxicity testing which is necessary to ensure that the risks associated with ophthalmic products meet appropriate safety criteria and are clearly labelled. In vitro preclinical testing is now a well-established practice of significant importance for evaluating the efficacy and safety of cosmetic, pharmaceutical, and nutraceutical products. Along with in vitro testing, also computational procedures, herein described, for evaluating the pharmacological profile of potential ocular drug candidates including their toxicity, are in rapid expansion. In this review the ocular cell types and functionality are described providing an overview about the scientific challenge for the development of three-dimensional in vitro models.

scholarly journals Development of In Vitro Corneal Models: Opportunity for Pharmacological Testing

2020 ◽  
Author(s):  
Valentina Citi ◽  
Eugenia Piragine ◽  
Simone Brogi ◽  
Sara Ottino ◽  
Vincenzo Calderone
Keyword(s):  
Three Dimensional ◽  
Toxicity Testing ◽  
Cell Types ◽  
In Vitro Models ◽  
Rapid Expansion ◽  
Anatomy And Physiology ◽  
Complex Anatomy ◽  
Computational Procedures ◽  
Scientific Challenge

Human eye is a specialized organ with complex anatomy and physiology, because it is characterized by different cell types with specific physiological functions. Given the complexity of the eye, ocular tissues are finely organized and orchestrated. In the last few years many in vitro models have been developed, in order to meet the 3Rs principle (Replacement, Reduction and Refinement) for eye toxicity testing. This procedure is highly necessary to ensure that the risks associated with ophthalmic products meet appropriate safety criteria. In vitro preclinical testing is now a well-established practice of significant importance for evaluating the efficacy and safety of cosmetic, pharmaceutical, and nutraceutical products. Along with in vitro testing, also computational procedures, herein described, for evaluating the pharmacological profile of potential ocular drug candidates including their toxicity, are in rapid expansion. In this review the ocular cell types and functionality are described providing an overview about the scientific challenge for the development of three-dimensional in vitro models.

scholarly journals Emerging Neuroblastoma 3D In Vitro Models for Pre-Clinical Assessments

2020 ◽  
Vol 11 ◽  
Author(s):  
Diana Corallo ◽  
Stella Frabetti ◽  
Olivia Candini ◽  
Elisa Gregianin ◽  
Massimo Dominici ◽  
...  
Keyword(s):  
Three Dimensional ◽  
Structural Complexity ◽  
3D Cell Culture ◽  
Cell Types ◽  
In Vitro Models ◽  
Cancer Therapies ◽  
Extracellular Matrix Components ◽  
Anti Cancer

The potential of tumor three-dimensional (3D) in vitro models for the validation of existing or novel anti-cancer therapies has been largely recognized. During the last decade, diverse in vitro 3D cell systems have been proposed as a bridging link between two-dimensional (2D) cell cultures and in vivo animal models, both considered gold standards in pre-clinical settings. The latest awareness about the power of tailored therapies and cell-based therapies in eradicating tumor cells raises the need for versatile 3D cell culture systems through which we might rapidly understand the specificity of promising anti-cancer approaches. Yet, a faithful reproduction of the complex tumor microenvironment is demanding as it implies a suitable organization of several cell types and extracellular matrix components. The proposed 3D tumor models discussed here are expected to offer the required structural complexity while also assuring cost-effectiveness during pre-selection of the most promising therapies. As neuroblastoma is an extremely heterogenous extracranial solid tumor, translation from 2D cultures into innovative 3D in vitro systems is particularly challenging. In recent years, the number of 3D in vitro models mimicking native neuroblastoma tumors has been rapidly increasing. However, in vitro platforms that efficiently sustain patient-derived tumor cell growth, thus allowing comprehensive drug discovery studies on tailored therapies, are still lacking. In this review, the latest neuroblastoma 3D in vitro models are presented and their applicability for a more accurate prediction of therapy outcomes is discussed.

Abstract 658: Unraveling the Controversy of Bisphosphonates as Vascular Calcification Therapy Using a Nanoanalytical Approach

2016 ◽  
Vol 36 (suppl_1) ◽  
Author(s):  
Jessica L Ruiz ◽  
Joshua D Hutcheson ◽  
Elena Aikawa
Keyword(s):  
Vascular Calcification ◽  
Plaque Rupture ◽  
Three Dimensional ◽  
Building Blocks ◽  
Underlying Mechanism ◽  
Pharmacological Intervention ◽  
Fibrous Cap ◽  
Collagen Hydrogel ◽  
Increased Risk

Vascular calcification significantly predicts atherosclerotic plaque rupture and cardiovascular events. Retrospective studies of women taking bisphosphonates, a proposed therapy for vascular calcification, paradoxically indicated increased risk in patients with prior acute events. We recently demonstrated that calcifying extracellular vesicles (EVs) released by cells within the plaque aggregate and nucleate calcific mineral, but the underlying mechanism and the potential for pharmacological intervention remain poorly understood. We hypothesize that bisphosphonates block EV aggregation and arrest existing mineral growth, freezing calcifications in a high-risk morphology that hastens plaque rupture. This study visualized for the first time EV aggregation and calcification at single-EV resolution, via scanning electron microscopy. Three-dimensional (3-D) collagen hydrogels incubated with calcifying EVs modeled fibrous cap calcification, serving as an in vitro platform to image mineral nucleation and test candidate drugs for the potential to inhibit or reverse vascular calcification. EVs aggregated along and between collagen fibrils. Energy-dispersive x-ray spectroscopy (EDS) confirmed that EV aggregates contained calcium and phosphorous, the building blocks of calcific mineral (vs. internal collagen control, p<0.001). The addition of the bisphosphonate ibandronate decreased the EDS-detected amount of calcium (4.32% by weight (wt%) vs. 2.36 wt%, p<0.001) and phosphorous (4.26 wt% vs. 1.94 wt%, p<0.001) comprising EV aggregates. Further, ibandronate reduced the size (21.5 μm 2 vs. 14.2 μm 2 , p=0.012) and changed the morphology of calcific EV aggregates (Figure). These findings agree with our hypothesis that bisphosphonates alter EV-driven calcification, and confirm that our 3-D collagen hydrogel system is a viable platform to study EV-mediated mineral nucleation and evaluate potential therapies for cardiovascular calcification.

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