scholarly journals Application of LDH assay for therapeutic efficacy evaluation of ex vivo tumor models

2021 ◽  
Vol 11 (1) ◽  
Author(s):  
Megan C. Cox ◽  
Rita Mendes ◽  
Fernanda Silva ◽  
Teresa F. Mendes ◽  
Adelyn Zelaya-Lazo ◽  
...  
Keyword(s):  
Therapeutic Efficacy ◽  
Ex Vivo ◽  
3D Culture ◽  
Attrition Rate ◽  
Added Value ◽  
Tumor Models ◽  
Efficacy Evaluation ◽  
Culture Systems ◽  
Ldh Assay ◽  
The Impact

AbstractThe current standard preclinical oncology models are not able to fully recapitulate therapeutic targets and clinically relevant disease biology, evidenced by the 90% attrition rate of new therapies in clinical trials. Three-dimensional (3D) culture systems have the potential to enhance the relevance of preclinical models. However, the limitations of currently available cellular assays to accurately evaluate therapeutic efficacy in these models are hindering their widespread adoption. We assessed the compatibility of the lactate dehydrogenase (LDH) assay in 3D spheroid cultures against other commercially available readout methods. We developed a standardized protocol to apply the LDH assay to ex vivo cultures, considering the impact of culture growth dynamics. We show that accounting for growth rates and background release levels of LDH are sufficient to make the LDH assay a suitable methodology for longitudinal monitoring and endpoint assessment of therapeutic efficacy in both cell line-derived xenografts (xenospheres) and patient-derived explant cultures. This method has the added value of being non-destructive and not dependent on reagent penetration or manipulation of the parent material. The establishment of reliable readout methods for complex 3D culture systems will further the utility of these tumor models in preclinical and co-clinical drug development studies.

scholarly journals Characterizing the impact of 2D and 3D culture conditions on the therapeutic effects of human mesenchymal stem cell secretome on corneal wound healing in vitro and ex vivo

2019 ◽  
Vol 99 ◽  
pp. 247-257 ◽  
Author(s):  
Kaylene Carter ◽  
Hyun Jong Lee ◽  
Kyung-Sun Na ◽  
Gabriella Maria Fernandes-Cunha ◽  
Ignacio Jesus Blanco ◽  
...  
Keyword(s):  
Wound Healing ◽  
Ex Vivo ◽  
3D Culture ◽  
Human Mesenchymal Stem Cell ◽  
Culture Conditions ◽  
Therapeutic Effects ◽  
Corneal Wound ◽  
2D And 3D ◽  
The Impact

scholarly journals Self-Assembling Peptide Hydrogels - PeptiGels® as a Platform for Hepatic Organoid Culture

2021 ◽  
Author(s):  
Adedamola Olayanju ◽  
Aline F Miller ◽  
Tahera Ansari ◽  
Christopher E. Goldring
Keyword(s):  
Ex Vivo ◽  
3D Culture ◽  
Molecular Techniques ◽  
3 Dimensional ◽  
Culture Systems ◽  
Self Assembling ◽  
Technological Platform ◽  
Peptide Hydrogels

AbstractA major challenge in advancing preclinical studies is the lack of robust in vitro culture systems that fully recapitulate the in vivo scenario together with limited clinical translational to humans. Organoids, as 3-dimensional (3D) self-replicating structures are increasingly being shown as powerful models for ex vivo experimentation in the field of regenerative medicine and drug discovery. Organoid formation requires the use of extracellular matrix (ECM) components to provide a 3D platform. However, the most commonly used ECM, essential for maintaining organoid growth is Matrigel and is derived from a tumorigenic source which limits its translational ability. PeptiGels® which are self-assembling peptide hydrogels present as alternatives to traditional ECM for use in 3D culture systems. Synthetic PeptiGels® are non-toxic, biocompatible, biodegradable and can be tuneable to simulate different tissue microenvironments. In this study, we validated the use of different types of PeptiGels® for porcine hepatic organoid growth. Hepatic organoids were assessed morphologically and using molecular techniques to determine the optimum PeptiGel® formulation. The outcome clearly demonstrated the ability of PeptiGel® to support organoid growth and offer themselves as a technological platform for 3D cultured physiologically and clinically relevant data.

scholarly journals From Clones to Buds and Branches: The Use of Lung Organoids to Model Branching Morphogenesis Ex Vivo

2021 ◽  
Vol 9 ◽  
Author(s):  
Ana Ivonne Vazquez-Armendariz ◽  
Susanne Herold
Keyword(s):  
Cell Fate ◽  
Molecular Mechanisms ◽  
Ex Vivo ◽  
3D Culture ◽  
Cell Types ◽  
Branching Morphogenesis ◽  
Cell Fate Decisions ◽  
Culture Systems

Three-dimensional (3D) organoid culture systems have rapidly emerged as powerful tools to study organ development and disease. The lung is a complex and highly specialized organ that comprises more than 40 cell types that offer several region-specific roles. During organogenesis, the lung goes through sequential and morphologically distinctive stages to assume its mature form, both structurally and functionally. As branching takes place, multipotent epithelial progenitors at the distal tips of the growing/bifurcating epithelial tubes progressively become lineage-restricted, giving rise to more differentiated and specialized cell types. Although many cellular and molecular mechanisms leading to branching morphogenesis have been explored, deeper understanding of biological processes governing cell-fate decisions and lung patterning is still needed. Given that these distinct processes cannot be easily analyzedin vivo, 3D culture systems have become a valuable platform to study organogenesisin vitro. This minireview focuses on the current lung organoid systems that recapitulate developmental events occurring before and during branching morphogenesis. In addition, we also discuss their limitations and future directions.

Use of 3D culture systems to generate human induced pluripotent stem cell-derived [beta]-cells in vitro

2018 ◽  
Author(s):  
Fantuzzi Federica ◽  
Toivonen Sanna ◽  
Schiavo Andrea Alex ◽  
Pachera Nathalie ◽  
Rajaei Bahareh ◽  
...  
Keyword(s):  
Stem Cell ◽  
Beta Cells ◽  
Pluripotent Stem Cell ◽  
3D Culture ◽  
Induced Pluripotent Stem Cell ◽  
Culture Systems ◽  
Induced Pluripotent

scholarly journals 231 Impact of Brachyspira hyodysentariae on intestinal function and integrity

2020 ◽  
Vol 98 (Supplement_3) ◽  
pp. 116-116
Author(s):  
Emma T Helm ◽  
Susanne J Lin ◽  
Nicholas Gabler ◽  
Eric R Burrough
Keyword(s):  
Ex Vivo ◽  
Potato Starch ◽  
Nutrient Digestibility ◽  
High Morbidity ◽  
Post Inoculation ◽  
Intestinal Function ◽  
Treatment Groups ◽  
Intestinal Integrity ◽  
Beet Pulp ◽  
The Impact

Abstract Swine dysentery (SD) induced by Brachyspira hyodysentariae (Bhyo) causes colitis and mucohemorrhagic diarrhea in grow-finish pigs, however little is known about the physiological changes that occur to the gastrointestinal tract during Bhyo infection. Thus, the objective of this study was to evaluate the impact of a Bhyo challenge on intestinal function and integrity of pigs fed two divergent diets. A total of 36 Bhyo negative gilts (24.3 ± 3.6 kg BW) were selected and assigned to one of three treatment groups (n=12 pigs/trt): 1) Bhyo negative, 20% DDGS diet (CON), 2) Bhyo challenged, 20% DDGS diet (DDGS), and 3) Bhyo challenged, 10% DDGS, 5% beet pulp and 5% resistant potato starch diet (RS). Pigs were fed diets 21 days prior to challenge and on days post inoculation (dpi) 0 and 1, pigs were inoculated with Bhyo or sham. Fecal samples were collected for ATTD and pigs were euthanized for colon collection within 72 hours of initial observation of clinical SD, or at the end of the study (dpi 10-16). Tissues were assessed for ex vivo measures of intestinal integrity and mitochondrial function. The challenge resulted in high morbidity, with 88% of DDGS and RS pigs developing clinical SD. Colon transepithelial resistance was increased in DDGS pigs compared with CON and RS pigs (P=0.005), and colon macromolecule permeability was reduced in both DDGS and RS pigs compared with CON pigs (P=0.006), likely due to mucoid discharge. Colonic mitochondrial oxygen consumption was not impacted by treatment (P >0.10). Further, ATTD of DM, OM, N, and GE were reduced in DDGS pigs compared with CON pigs (P< 0.001), whilst nutrient digestibility was not reduced in RS pigs. Taken together, these data show Bhyo does not appear to reduce ex vivo colonic integrity. Further, the RS diet may reduce severity of a Bhyo challenge.

scholarly journals SUPPORT FOR MANAGING PARTLY CONFIGURABLE MODULAR SYSTEMS

2021 ◽  
Vol 1 ◽  
pp. 2791-2800
Author(s):  
Jarkko Pakkanen ◽  
Teuvo Heikkinen ◽  
Nillo Adlin ◽  
Timo Lehtonen ◽  
Janne Mämmelä ◽  
...  
Keyword(s):  
Portfolio Management ◽  
Product Variety ◽  
Added Value ◽  
Modular System ◽  
Product Management ◽  
Design Decision ◽  
Product Portfolio ◽  
Modular Systems ◽  
Engineer To Order ◽  
The Impact

AbstractThe paper studies what kind of support could be applied to the management of partly configurable modular systems. The main tasks of product management, product portfolio management and product variety management are defined. In addition, a partly configurable product structure and modular system are defined. Because the limited support in the literature for managing partly configurable modular systems, the article reviews previous product development cases in which authors have been involved on lessons learnt basis, i.e., if the methods and tools used in the cases could provide support for the research objective. As a result, the existing definition of the modular system should be extended by the concepts of non-module and design decision sequence description when dealing with partly configurable modular systems. This is because engineer-to-order should be made possible in cases where it brings clear added value to the customer compared to completely pre-defined solutions that may limit the customer's interest in the offering. Tools to assess the impact of changes to the product offering are required. These should be taken into account in frameworks that are used in method and tool development.

scholarly journals Histological, Biomechanical, and Biological Properties of Genipin-Crosslinked Decellularized Peripheral Nerves

2021 ◽  
Vol 22 (2) ◽  
pp. 674
Author(s):  
Óscar Darío García-García ◽  
Marwa El Soury ◽  
David González-Quevedo ◽  
David Sánchez-Porras ◽  
Jesús Chato-Astrain ◽  
...  
Keyword(s):  
Ex Vivo ◽  
Nerve Repair ◽  
Wistar Rat ◽  
Biomechanical Properties ◽  
Biological Properties ◽  
Suitable Alternative ◽  
Promising Alternative ◽  
Histological Pattern ◽  
The Impact

Acellular nerve allografts (ANGs) represent a promising alternative in nerve repair. Our aim is to improve the structural and biomechanical properties of biocompatible Sondell (SD) and Roosens (RS) based ANGs using genipin (GP) as a crosslinker agent ex vivo. The impact of two concentrations of GP (0.10% and 0.25%) on Wistar rat sciatic nerve-derived ANGs was assessed at the histological, biomechanical, and biocompatibility levels. Histology confirmed the differences between SD and RS procedures, but not remarkable changes were induced by GP, which helped to preserve the nerve histological pattern. Tensile test revealed that GP enhanced the biomechanical properties of SD and RS ANGs, being the crosslinked RS ANGs more comparable to the native nerves used as control. The evaluation of the ANGs biocompatibility conducted with adipose-derived mesenchymal stem cells cultured within the ANGs confirmed a high degree of biocompatibility in all ANGs, especially in RS and RS-GP 0.10% ANGs. Finally, this study demonstrates that the use of GP could be an efficient alternative to improve the biomechanical properties of ANGs with a slight impact on the biocompatibility and histological pattern. For these reasons, we hypothesize that our novel crosslinked ANGs could be a suitable alternative for future in vivo preclinical studies.

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