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Cancer Research
在斑马鱼胚胎中使用白血病细胞移植 物进行体内 药物反应的快速预测
在斑马鱼胚胎中使用白血病细胞移植 物进行体内 药物反应的快速预测
JoVE Journal
Cancer Research
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JoVE Journal Cancer Research
Rapid in vivo Drug Response Prediction Using Leukemia Cell Grafts in Zebrafish Embryos

在斑马鱼胚胎中使用白血病细胞移植 物进行体内 药物反应的快速预测

Full Text
1,029 Views
10:46 min
May 23, 2025

DOI: 10.3791/67451-v

Sophia Sartorius1, Luca Bramè1,2, Jutta Proba1, Anton Gauert1, Nadine Olk1, Juan Lazaro1, Angelika Eggert1, Cornelia Eckert1, Anja H. Hagemann1,2

1Charité-Universitätsmedizin Berlin, Klinik für Pädiatrie m.S. Onkologie/Hämatologie, 2German Cancer Consortium (DKTK), partner site Berlin, and German Cancer Research Center (DKFZ)

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Overview

This protocol outlines a rapid seven-day workflow for assessing drug responses in human acute lymphoblastic leukemia (ALL) using zebrafish embryos. It details the generation of xenografts from patient-derived leukemia cells and provides insights into adapting the method for solid tumors.

Key Study Components

Area of Science

  • Oncology
  • Pharmacology
  • Preclinical Models

Background

  • Human acute lymphoblastic leukemia (ALL) is a pediatric cancer.
  • Zebrafish embryos serve as a unique preclinical model for studying cancer.
  • Access to rare cancer samples enhances the relevance of the study.
  • Drug penetration and pharmacokinetics are critical for effective treatment assessment.

Purpose of Study

  • To develop a protocol for generating xenografts from ALL and solid tumors.
  • To evaluate drug responses in a clinically relevant model.
  • To improve the survival of graft cells by blocking immune differentiation.

Methods Used

  • Generation of xenografts from cell lines and fresh patient material.
  • Transient immunosuppression of zebrafish embryos.
  • Flow cytometry for assessing single cell viability and proliferation.
  • Testing drug penetration before experimental trials.

Main Results

  • Successful growth of patient-derived leukemia cells in zebrafish.
  • Adaptation of the protocol for solid pediatric tumors.
  • Effective measurement of drug responses using flow cytometry.
  • Improved graft cell survival through immune cell differentiation blockade.

Conclusions

  • The zebrafish model is a promising tool for pediatric oncology research.
  • Drug response assessment can be effectively conducted in this model.
  • Future studies may expand to other types of pediatric tumors.

Frequently Asked Questions

What types of cancer can be studied using this protocol?
This protocol is designed for human acute lymphoblastic leukemia (ALL) and can be adapted for solid pediatric tumors.
How does the zebrafish model benefit cancer research?
Zebrafish embryos provide a unique preclinical model that allows for the study of drug responses in a living organism.
What is the significance of using flow cytometry in this study?
Flow cytometry allows for precise measurement of single cell viability and proliferation, which is crucial for assessing drug responses.
Why is pharmacokinetics important in this research?
Understanding pharmacokinetics ensures that drugs can effectively penetrate biological barriers to reach the target cells.
What is the duration of the experimental workflow?
The experimental workflow is designed to be completed in seven days.
How does the protocol address immune cell differentiation?
The protocol includes methods to block innate immune cell differentiation, enhancing the survival of graft cells.

该方案提供了从细胞系和瞬时免疫抑制斑马鱼胚胎中的新鲜患者材料生成和排除人急性淋巴细胞白血病 (ALL) 异种移植物的分步说明,以及使用流式细胞术评估药物反应的指南。实验管道也可以适用于实体瘤。

我们开发了一种快速的 7 天工作流程,以测试肿瘤细胞对使用斑马鱼胚胎的药物的反应。由于靠近儿科肿瘤学,我们可以获得罕见的癌症样本,我们从 B 细胞前体白血病开始,但我们现在正在采用该方案来治疗儿科实体瘤,看看该检测是否也适用于临床环境。我们可以在斑马鱼胚胎中成功培养患者来源的白血病细胞,这使它们成为一种了不起的临床前模型。

然而,药代动力学可能很棘手,因为有些药物难以穿过生物屏障,因此我们确保在开始实验之前测试渗透性。我们的方法很独特,因为它阻断了先天免疫细胞分化,这有助于白血病的移植细胞更好地生存。此外,我们正在使用流式细胞术来测量 10 条宿主鱼池中移植细胞的单细胞活力和增殖。

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