This document describes the use of 3D neurosphere assays to study developmental neurotoxicity. Neural stem/progenitor cells from fetal rat and human brains are cultured as neurospheres to model processes of brain development in vitro. The neurosphere assay can assess effects of compounds on proliferation, migration, differentiation and other pathways. It allows medium-throughput testing of compounds using automated imaging and analysis. Species-specific effects of compounds like valproic acid on rat and human neurospheres have been observed. The neurosphere assay is useful for safety testing and drug development by providing a 3D in vitro model of the developing brain.
Microdialysis is an integral part of preclinical research to determine extracellular fluid and blood concentrations of metabolites, hormones, drugs, etc, and is often used in quantifying the biochemistry of brain and peripheral tissues. However, it is a molecular-only technique and other imaging modalities are needed to provide the researcher with functional and anatomical information of the animal in vivo.
Compare density gradient centrifugation, Magnet-activated cell sorting (MACS), and Fluorescence-activated cell sorting (FACS) in the isolation of pure stem cell populations from a heterogeneous suspension. Discuss the advantages and disadvantages of each technology as well as the emerging (recent) methods in stem cells separation.
This paper resulted from joint research between Thai and Oz researchers at the Australian Synchrotron in 2008. It represents a on-going collaboration between Siam Photon, Suranaree University of Technology, Monash University and the Australian Synchrotron
Microdialysis is an integral part of preclinical research to determine extracellular fluid and blood concentrations of metabolites, hormones, drugs, etc, and is often used in quantifying the biochemistry of brain and peripheral tissues. However, it is a molecular-only technique and other imaging modalities are needed to provide the researcher with functional and anatomical information of the animal in vivo.
Compare density gradient centrifugation, Magnet-activated cell sorting (MACS), and Fluorescence-activated cell sorting (FACS) in the isolation of pure stem cell populations from a heterogeneous suspension. Discuss the advantages and disadvantages of each technology as well as the emerging (recent) methods in stem cells separation.
This paper resulted from joint research between Thai and Oz researchers at the Australian Synchrotron in 2008. It represents a on-going collaboration between Siam Photon, Suranaree University of Technology, Monash University and the Australian Synchrotron
Medcrave Group - Microfluidic technologiesMedCrave
Exosomes are cell-released small membrane vesicles derived from the endolysosomal pathway with a size range of 30-150 nm. Since the first discovery in 1981, exosomes have been found to be released from various cell types and present in many biological fluids, including blood, urine, erebrospinal fluid and ascites. Significant attention has been focused on exosome molecular components (e.g. roteins, mRNA and miRNA) which have been implicated in a variety of physiological functions and pathological disease states.
Nanotechnology and its Application in Cancer TreatmentHasnat Tariq
Nanotechnology
Nanomaterials
Nanostructures
Nanoparticles
Unexpected Optical Properties of Nanoparticles
Synthesis of Nanoparticles
Nanotechnology in Cancer Treatment
Role of Sulfur NPs in Cancer Treatment
Human Tumour Cell Lines Used in Research
Ehrlich ascites carcinoma (EAC)
Sulfur Nanoparticles Preparation
MTT Assay
Sulphorhodamine-B (SRB) Assay
Median lethal dose (LD 50)
Experimental design
FT-IR Characterization of Sulfur Nanoparticles
SEM Characterization of Sulfur Nanoparticles
EDS Characterization of Sulfur Nanoparticles
XRD Characterization of Sulfur Nanoparticles
Chemical Studies on Sulfur Nanoparticles In Vitro
Biochemical investigations
Conclusion
Applications of Nanoparticles in cancer treatment
Nanoshells
Nano X-Ray therapy
Drug Delivery by Nanoparticles
Anticancer drug discovery using multicellular tumor spheroid modelsHasnat Tariq
Cancer, drug discovery, tumor spheroids, organoids, 3D tumor spheroids, 3D scaffold-based models, Scaffold-free models, 3D Scaffolds, Hanging drop, Low adhesion microplate, Magnetic levitation and bio printing, bioprinting, anticancer,, tumor models, Drug screening assays, flow cytometry, expansion microscopy.
Medcrave Group - Microfluidic technologiesMedCrave
Exosomes are cell-released small membrane vesicles derived from the endolysosomal pathway with a size range of 30-150 nm. Since the first discovery in 1981, exosomes have been found to be released from various cell types and present in many biological fluids, including blood, urine, erebrospinal fluid and ascites. Significant attention has been focused on exosome molecular components (e.g. roteins, mRNA and miRNA) which have been implicated in a variety of physiological functions and pathological disease states.
Nanotechnology and its Application in Cancer TreatmentHasnat Tariq
Nanotechnology
Nanomaterials
Nanostructures
Nanoparticles
Unexpected Optical Properties of Nanoparticles
Synthesis of Nanoparticles
Nanotechnology in Cancer Treatment
Role of Sulfur NPs in Cancer Treatment
Human Tumour Cell Lines Used in Research
Ehrlich ascites carcinoma (EAC)
Sulfur Nanoparticles Preparation
MTT Assay
Sulphorhodamine-B (SRB) Assay
Median lethal dose (LD 50)
Experimental design
FT-IR Characterization of Sulfur Nanoparticles
SEM Characterization of Sulfur Nanoparticles
EDS Characterization of Sulfur Nanoparticles
XRD Characterization of Sulfur Nanoparticles
Chemical Studies on Sulfur Nanoparticles In Vitro
Biochemical investigations
Conclusion
Applications of Nanoparticles in cancer treatment
Nanoshells
Nano X-Ray therapy
Drug Delivery by Nanoparticles
Anticancer drug discovery using multicellular tumor spheroid modelsHasnat Tariq
Cancer, drug discovery, tumor spheroids, organoids, 3D tumor spheroids, 3D scaffold-based models, Scaffold-free models, 3D Scaffolds, Hanging drop, Low adhesion microplate, Magnetic levitation and bio printing, bioprinting, anticancer,, tumor models, Drug screening assays, flow cytometry, expansion microscopy.
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As shown by AstraZeneca in nature reviews*, one third of safety failures along the drug discovery process is linked to CNS toxicity uncovered in clinical trials. To avoid this attrition, the potential neurotoxicity of any drug going through the blood brain barrier (BBB) needs to be assessed in the very early stages of new chemical entities (NCE) research. Neurotoxicity assays can be performed on the SH-SY5Y human cell line by using High-Content Screening (HCS) technologies. The present study was performed using classical 2D and 3D culture protocols. In this poster, 2D results and preliminary 3D culture results on multiple reference compounds are depicted.
In Vitro Neuroscience Services-Creative Biolabscailynnjohnson
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https://neuros.creative-biolabs.com/
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MINI REVIEW ARTICLEpublished 04 March 2015doi 10.3389.docxannandleola
MINI REVIEW ARTICLE
published: 04 March 2015
doi: 10.3389/fnins.2015.00064
Lessons from the analysis of nonhuman primates for
understanding human aging and neurodegenerative
diseases
Jean-Michel Verdier 1,2,3*, Isabelle Acquatella 1,2,3, Corinne Lautier 1,2,3, Gina Devau 1,2,3,
Stéphanie Trouche 1,2,3, Christelle Lasbleiz 1,2,3 and Nadine Mestre-Francés 1,2,3
1 Université de Montpellier, Montpellier, France
2 Institut National de la Santé et de la Recherche Médicale, U1198, Montpellier, France
3 Ecole Pratique des Hautes Etudes, Paris, France
Edited by:
Patrick Lewis, University of Reading,
UK
Reviewed by:
Ricardo Insausti, University of
Castilla -la Mancha, Spain
Varun Kesherwani, University of
Nebraska Medical Center, USA
*Correspondence:
Jean-Michel Verdier, University of
Montpellier, Place Eugène Bataillon,
CC105, 34095 Montpellier, France
e-mail: [email protected]
univ-montp2.fr
Animal models are necessary tools for solving the most serious challenges facing medical
research. In aging and neurodegenerative disease studies, rodents occupy a place of
choice. However, the most challenging questions about longevity, the complexity and
functioning of brain networks or social intelligence can almost only be investigated in
nonhuman primates. Beside the fact that their brain structure is much closer to that of
humans, they develop highly complex cognitive strategies and they are visually-oriented
like humans. For these reasons, they deserve consideration, although their management
and care are more complicated and the related costs much higher. Despite these caveats,
considerable scientific advances have been possible using nonhuman primates. This
review concisely summarizes their role in the study of aging and of the mechanisms
involved in neurodegenerative disorders associated mainly with cognitive dysfunctions
(Alzheimer’s and prion diseases) or motor deficits (Parkinson’s and related diseases).
Keywords: neuroscience, brain, aging, neurodegenerative diseases, nonhuman primate
WHY DO WE NEED ANIMAL MODELS?
The simplest answer to this question is to increase our general
knowledge, to experimentally test theories. Animal model use-
fulness is manifold, from the study of physiological processes to
the identification of disease-causing mechanisms. Indeed, phys-
iopathological studies are of the utmost importance for develop-
ing diagnostic and therapeutic approaches based on the discovery
of new, more sensitive and specific biomarkers, the identifica-
tion of the mechanism of action of drugs, the establishment of
pharmacodynamics and pharmacokinetic parameters, the toxic-
ity analysis of new compounds or the assessment of clinical drug
regimens.
Many different animal models, ranging from unicellular
organisms (bacteria, yeast) to invertebrates (the roundworm
Caenorhabditis elegans or the fruit fly Drosophila melanogaster)
and vertebrates (fish and mammals), are currently used for
research on aging and neurodegenerative disorders. T ...
This webinar is part of a 2-hour monthly series hosted by the Neurotechnology Innovation Network: https://ktn-uk.org/health/neurotechnology/
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The in vitro choice ellen iuf 2013
1. NEUROSPHERES AS
3D IN VITRO MODELS
FOR ASSESSING DEVELOPMENTAL
NEUROTOXICITY
Prof. Ellen Fritsche
IUF- Leibniz Research Institute for Environmental Medicine
13. The ‘Neurosphere-Assay’
Apoptosis
Fritsche et al. Environ Health Perspect 2005
Moors et al. Toxicol Appl Pharmacol 2007
Moors et al. Environ Health Perspect 2009
Moors et al. Genes & Immunity 2010
Tegenge et al. Cell. Mol. Life Sci. 2010
Schreiber et al. Environ Health Perspect 2010
Gassmann et al. Environ Health Perspect 2010
Verner et al. Toxicol in Vitro 2011
BrdU
Fritsche et al. Methods Mol Biol 2011
Gassmann et al. Toxicol in Vitro 2012
Bal-Price et al. ALTEX 2012
From Breier…Fritsche.. et al. Neurotoxicol Teratol 2009
14. The Neurosphere Method in Medium Throughput
Medium Throughput:
• Migration
• Neuronal Differentiation
• Neuronal Migration
Cellomics ArrayScan:
Nuclei and beta(III)tubulin
stainings of a differentiated
Human Neurosphere
15. Summary & Conclusion
• Neurospheres of different species mimic processes of brain development in vitro.
• Application domains of neurospheres include safety testing and drug
development.
• Species-specific VPA effects on rat and human neurospheres.
• Medium throughput is possible for compound testing in human and rodent
neurospheres.
The 3D neurosphere assay can be used to study species-specific pathway-to-function
effects (positive/negative) of compounds in a medium throughput fashion by
automated plating and usage of ‘High Content Image Analyses’.
16. Acknowledgements
Dr. Marta Barenys
Dr. Kathrin Gassmann
Dr. Julia Tigges
Dr. Susanne Giersiefer
Dr. Janette Schuwald
Jenny Baumann
Katharina Dach
Martin Schmuck
Maxi Hofrichter
Katerina Kefalaki
Stefan Masjosthusmann
Thomas Temme
Denise de Boer
Ulrike Hübenthal
Cooperations:
High Content Imaging: Axel Mosig, University of Bochum
Proteomics: Prof. Kai Stühler, HHU Düsseldorf
LRI Innovative
Science Award 2006
German Alternative
Methods Award 2007
Leibniz DAAD
Thank you for your attention!