Drug Screening of Primary Patient Derived Tumor Xenografts in Zebrafish

Haney, Meghan G.; Moore, L. Henry; Blackburn, Jessica S.

doi:10.3791/60996

Cited by 14 publications

(17 citation statements)

References 18 publications

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“…The evaluation of cell viability using DiI labeling in zebrafish xenograft models has been widely used for different cancer cell models, such as melanoma cells [ 31 ] and leukemia cells [ 32 ]. Furthermore, this labeling can be applied for high-throughput drug screening by incorporating an automatic fluorescent microscopy system [ 33 , 34 ].…”

Section: Resultsmentioning

confidence: 99%

Microcrater-Arrayed Chemiluminescence Cell Chip to Boost Anti-Cancer Drug Administration in Zebrafish Tumor Xenograft Model

Kuo

Lai

et al. 2021

Biology

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Purpose: The aim of this study was to develop a rapid and automatic drug screening platform using microcrater-arrayed (µCA) cell chips. Methods: The µCA chip was fabricated using a laser direct writing technique. The fabrication time required for one 9 × 9 microarray wax chip was as quick as 1 min. On a nanodroplet handling platform, the chip was pre-coated with anti-cancer drugs, including cyclophosphamide, cisplatin, doxorubicin, oncovin, etoposide, and 5-fluorouracil, and their associated mixtures. Cell droplets containing 100 SK-N-DZ or MCF-7 cells were then loaded onto the chip. Cell viability was examined directly through a chemiluminescence assay on the chip using the CellTiter-Glo assay. Results: The time needed for the drug screening assay was demonstrated to be less than 30 s for a total of 81 tests. The prediction of optimal drug synergy from the µCA chip was found by matching it to that of the zebrafish MCF-7 tumor xenograft model, instead of the conventional 96-well plate assay. In addition, the critical reagent volume and cell number for each µCA chip test were 200 nL and 100 cells, respectively, which were significantly lower than 100 µL and 4000 cells, which were achieved using the 96-well assay. Conclusion: Our study for the µCA chip platform could improve the high-throughput drug synergy screening targeting the applications of tumor cell biology.

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Section: Resultsmentioning

confidence: 99%

Microcrater-Arrayed Chemiluminescence Cell Chip to Boost Anti-Cancer Drug Administration in Zebrafish Tumor Xenograft Model

Kuo

Lai

et al. 2021

Biology

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“… Pause point: Once images have been taken, quantification can be done at any point. Quantify fluorescence using FIJI Image J software and macro as previously described in Haney et al. 2020 .…”

Section: Step-by-step Methods Detailsmentioning

confidence: 99%

“…Quantify fluorescence using FIJI Image J software and macro as previously described in Haney et al. 2020 .…”

Section: Step-by-step Methods Detailsmentioning

confidence: 99%

Protocol for rapid assessment of the efficacy of novel Wnt inhibitors using zebrafish models

et al. 2021

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Summary Dysregulation of Wnt signaling is a hallmark of many cancers, and the development of effective, non-toxic small-molecule Wnt inhibitors is desirable. Off-target toxicities of new compounds are typically tested in mouse models, which is both costly and time consuming. Here, we present a rapid and inexpensive protocol to determine the in vivo toxicity and efficacy of novel Wnt inhibitors in zebrafish using a combination of a fluorescence reporter assay as well as eye rescue and fin regeneration assays. These experiments are completed within 1 week to rapidly narrow drug candidates before moving to more expensive pre-clinical testing. For complete details on the use and execution of this protocol, please refer to Zhang et al. (2020) .

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“…Several features of zebrafish larvae make them uniquely suited for these studies, including: (i) transparent bodies that allow for easy tumor cell injection; (ii) ability to use trackable fluorescent-tagged cells following transplantation; (iii) an immature immune system during early embryonic development, which reduces the chance of the immune rejection of transplanted tumor cells; and (iv) availability of large clutches of embryos for transplantation. Multiple injection sites, such as the perivitelline space, pericardial space, yolk, retro-optical region, and brain, have been explored in a variety of studies to understand the mechanisms of tumor metastasis, angiogenesis, cellular intravasation/extravasation [ 90 , 91 , 92 ].…”

Section: Advantages Of Using Zebrafish As a Model For Nb Researchmentioning

confidence: 99%

Zebrafish as a Neuroblastoma Model: Progress Made, Promise for the Future

Yeo

Levee

et al. 2021

Cells

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For nearly a decade, researchers in the field of pediatric oncology have been using zebrafish as a model for understanding the contributions of genetic alternations to the pathogenesis of neuroblastoma (NB), and exploring the molecular and cellular mechanisms that underlie neuroblastoma initiation and metastasis. In this review, we will enumerate and illustrate the key advantages of using the zebrafish model in NB research, which allows researchers to: monitor tumor development in real-time; robustly manipulate gene expression (either transiently or stably); rapidly evaluate the cooperative interactions of multiple genetic alterations to disease pathogenesis; and provide a highly efficient and low-cost methodology to screen for effective pharmaceutical interventions (both alone and in combination with one another). This review will then list some of the common challenges of using the zebrafish model and provide strategies for overcoming these difficulties. We have also included visual diagram and figures to illustrate the workflow of cancer model development in zebrafish and provide a summary comparison of commonly used animal models in cancer research, as well as key findings of cooperative contributions between MYCN and diverse singling pathways in NB pathogenesis.

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Drug Screening of Primary Patient Derived Tumor Xenografts in Zebrafish

Cited by 14 publications

References 18 publications

Microcrater-Arrayed Chemiluminescence Cell Chip to Boost Anti-Cancer Drug Administration in Zebrafish Tumor Xenograft Model

Microcrater-Arrayed Chemiluminescence Cell Chip to Boost Anti-Cancer Drug Administration in Zebrafish Tumor Xenograft Model

Protocol for rapid assessment of the efficacy of novel Wnt inhibitors using zebrafish models

Zebrafish as a Neuroblastoma Model: Progress Made, Promise for the Future

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