Automated microinjection for zebrafish xenograft models

npj Biomedical Innovations., Journal Year: 2025, Volume and Issue: 2(1)

Published: April 23, 2025

Abstract Zebrafish xenograft models are increasingly recognized for predicting patient responses to cancer therapeutics, suggesting their potential as clinical diagnostic tools. However, precise microinjection of cells into numerous small and fragile zebrafish larvae is laborious, requires extensive training new operators, often yields variable results, limiting drug discovery applications. To address these challenges, we have designed, built, validated an automated robot. The robot performs injections the vasculature, perivitelline space, hindbrain ventricle in both fully semi-automated modes. Combined results demonstrate average injection success rate approximately 60% survival exceeding 70%, comparable manual methods, with mode being twice fast. This automation reduces need personnel while enhancing reproducibility, efficiency, accuracy, paving way more use diagnostics.

Language: Английский

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Speeding up Glioblastoma Cancer Research: Highlighting the Zebrafish Xenograft Model

International Journal of Molecular Sciences, Journal Year: 2024, Volume and Issue: 25(10), P. 5394 - 5394

Published: May 15, 2024

Glioblastoma multiforme (GBM) is a very aggressive and lethal primary brain cancer in adults. The multifaceted nature of GBM pathogenesis, rising from complex interactions between cells the tumor microenvironment (TME), has posed great treatment challenges. Despite significant scientific efforts, prognosis for remains poor, even after intensive with surgery, radiation, chemotherapy. Efficient management still requires invention innovative strategies. There strong necessity to complete vitro studies vivo properly evaluate mechanisms progression within TME. In recent years, animal models used study tumors have evolved, achieving highly invasive able provide key information on molecular onset. At present, most commonly research are represented by mammalian models, such as mouse canine ones. However, latter present several limitations, high cost time-consuming management, making them inappropriate large-scale anticancer drug evaluation. zebrafish (Danio rerio) model emerged valuable tool studying GBM. It shown promise preclinical due numerous advantages, its small size, ability generate large cohort genetically identical offspring, rapid development, permitting more time- cost-effective high-throughput screening when compared models. Moreover, transparent early developmental stages genetic anatomical similarities humans, it allows translatable related discovery. For this reason, aim review highlight potential relevant transgenic xenograft compare traditionally research.

Language: Английский

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Electrophysiological Impact of SARS-CoV-2 Envelope Protein in U251 Human Glioblastoma Cells: Possible Implications in Gliomagenesis?

International Journal of Molecular Sciences, Journal Year: 2024, Volume and Issue: 25(12), P. 6669 - 6669

Published: June 18, 2024

SARS-CoV-2 is the causative agent of COVID-19 pandemic, acute respiratory disease which, so far, has led to over 7 million deaths. There are several symptoms associated with infections which include neurological and psychiatric disorders, at least in case pre-Omicron variants. infection can also promote onset glioblastoma patients without prior malignancies. In this study, we focused on Envelope protein codified by virus genome, acts as viroporin that reported be central for propagation. particular, characterized electrophysiological profile E-protein transfected U251 HEK293 cells through patch-clamp technique FURA-2 measurements. Specifically, observed an increase voltage-dependent (Kv) calcium-dependent (KCa) potassium currents cell lines, respectively. Interestingly, both cellular models, a depolarization mitochondrial membrane potential accordance alteration growth. We, therefore, investigated transcriptional effect E signaling pathways found gene alterations apoptosis, cytokines WNT pathways. The changes after expression could explain impact gliomagenesis.

Language: Английский

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Uveal Melanoma Zebrafish Xenograft Models Illustrate the Mutation Status-Dependent Effect of Compound Synergism or Antagonism

Investigative Ophthalmology & Visual Science, Journal Year: 2024, Volume and Issue: 65(10), P. 26 - 26

Published: Aug. 20, 2024

Uveal melanoma (UM) is the most common primary intraocular malignancy with a high probability of metastatic disease. Although excellent treatment options for UM are available, therapy disease remain limited. Drug discovery studies using mouse models have thus far failed to provide therapeutic solutions, highlighting need novel models. Here, we optimize zebrafish xenografts as potential model drug by showcasing behavior multiple cell lines and findings on mutation-dependent compound synergism/antagonism Z-Tada; an algorithm objectively characterize output measurements.

Language: Английский

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Inhibition of GPR68 kills glioblastoma in zebrafish xenograft models

BMC Research Notes, Journal Year: 2024, Volume and Issue: 17(1)

Published: Aug. 23, 2024

Inhibition and knockdown of GPR68 negatively affects glioblastoma cell survival in vitro by inducing ferroptosis. Herein, we aimed to demonstrate that inhibition reduces the cells vivo using two orthotopic larval xenograft models Danio rerio, GBM lines U87-MG U138-MG. In cancer was assessed setting or knockdown. vitro, shRNA-mediated demonstrated potent cytotoxic effects against U87 U138 lines. This effect associated with increased intracellular lipid peroxidation, suggesting ferroptosis as underlying mechanism death. Translating these findings vivo, established a novel model zebrafish successfully grafting fluorescently labeled human cells, which were previously shown overexpress GPR68. shRNA significantly reduced viability grafted within this model. Additionally, treatment ogremorphin (OGM), highly specific small molecule inhibitor GPR68, also limited toxicity developing embryos. study suggests therapeutic targeting molecules like OGM represents promising approach for GBM.

Language: Английский

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MANEJO DAS DERMATOPATIAS NA ATENÇÃO PRIMÁRIA À SAÚDE

Published: June 11, 2024

The multidisciplinary approach in the management of polytrauma patients is utmost importance for proper evaluation and treatment injuries different body systems, as second most common cause death these occurs due to treatable injuries.The aim this article highlight correct through a review current literature.In article, we present literature widely used systematic approach.We discuss ABCDE sequence ATLS protocol, prioritizing airway control, breathing assessment, circulation hemorrhage neurological patient exposure injury identification.After stabilization identified injuries, detailed secondary assessment crucial.The emphasizes specialized team provide care optimize outcomes complex clinical situation.

Language: Английский

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AVANÇOS NA ABORDAGEM INICIAL AO POLITRAUMA

Published: June 11, 2024

The multidisciplinary approach in the management of polytrauma patients is utmost importance for proper evaluation and treatment injuries different body systems, as second most common cause death these occurs due to treatable injuries.The aim this article highlight correct through a review current literature.In article, we present literature widely used systematic approach.We discuss ABCDE sequence ATLS protocol, prioritizing airway control, breathing assessment, circulation hemorrhage neurological patient exposure injury identification.After stabilization identified injuries, detailed secondary assessment crucial.The emphasizes specialized team provide care optimize outcomes complex clinical situation.

Language: Английский

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An overview of glioblastoma multiforme <i>in vitro</i> experimental models

Journal of Biological Research - Bollettino della Società Italiana di Biologia Sperimentale, Journal Year: 2024, Volume and Issue: unknown

Published: Feb. 15, 2024

Glioblastoma multiforme (GBM) is the most common primary brain tumor, characterized by a remarkable inner complexity and inter-tumor variability. Moreover, it very aggressive resistant to conventional treatments, so that rapidly relapse. Therefore, there an immediate need for experimental strategies enhance our comprehension of GBM, aiming mitigate its economic social impact. Here, we described different in vivo vitro currently used study GBM. First, gave brief general overview classical models, including xenograft mouse zebrafish models canine offering wide range advantages but also presenting series strong limitations. Thus, starting from more traditional 2D culture comparing approaches critically exposing disadvantages using one or other methods. We briefly GBM systems allow recreating multiple cell-cell cell-extracellular matrix contacts still do not reflect tumors. finally intricacies novel 3D e.g., spheroids organoids. These sophisticated have demonstrated exceptional suitability across spectrum applications cancer research, ranging fundamental scientific inquiries translational research. Their adaptability three-dimensional architecture render them invaluable tools, new insights paving way advancements both basic applied

Language: Английский

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Uveal Melanoma zebrafish xenograft models illustrate the mutation status-dependent effect of compound synergism or antagonism

Research Square (Research Square), Journal Year: 2024, Volume and Issue: unknown

Published: April 24, 2024

Purpose Uveal melanoma (UM) is the most common primary intraocular malignancy with a high probability of metastatic disease. Although excellent treatment option for UM are available, therapy disease remain limited. Drug discovery studies using mouse models have thus far failed to provide therapeutic solutions, highlighting need novel models. Here, we optimize zebrafish xenografts as potential model drug by showcasing behavior multiple cell lines and findings on mutation-dependent compound synergism/antagonism Z-Tada; an algorithm objectively characterize output measurements. Methods Prognostic relevant or healthy melanocytes were inoculated at three distinct inoculation sites. Standardized quantifications independent site obtained measure tumor burden number, size distance disseminated cells. Sequentially, utilized this validate combinatorial synergism antagonism seen in vitro. Results Detailed analysis 691 demonstrated perivitelline space provided robust data dissemination. Cell more invasive (SF3B1^mut BAP1^mut) behaved aggressive in model. Combinatorial illustrated mutation-dependent, which confirmed vivo. differed per xenograft-model, it either inhibited overall Conclusion Perivitelline provides ability high-throughput screening acquisition Z-Tada. This demonstrates that uveal must take mutational subclasses into account, especially discoveries.

Language: Английский

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Glial plasticity in the zebrafish central nervous system

Trends in Cell Biology, Journal Year: 2024, Volume and Issue: 34(7), P. 531 - 534

Published: May 16, 2024

Glial cells have a remarkable plasticity. Recent studies using zebrafish as model highlight conserved cellular behavior in health and disease the central nervous system (CNS) between humans. These findings inform our understanding of their function how dysregulation pathogenesis can be determinant.

Language: Английский

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