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在本科生物学实验室中使用ibidi培养板介绍伤口愈合分析方法。

Introducing Wound Healing Assays in the Undergraduate Biology Laboratory Using Ibidi Plates.

作者信息

Suh Yewseok K, Robinson Ashley, Zanghi Nicholas, Kratz Austin, Gustetic Andrew, Crow Mackenzie M, Ritts Taylor, Hankey William, Segarra Verónica A

机构信息

Department of Biology, High Point University, High Point, North Carolina, USA.

Department of Chemistry, High Point University, High Point, North Carolina, USA.

出版信息

J Microbiol Biol Educ. 2022 Aug 8;23(2). doi: 10.1128/jmbe.00061-22. eCollection 2022 Aug.

Abstract

The wound healing assay is a simple and inexpensive method that allows researchers to experimentally mimic cell growth and migration leading to wound healing. In this assay, a wound is created on a monolayer of cultured mammalian cells and cell migration is monitored. Micrographs are captured at regular intervals during the duration of the experiment. These microscopy images are analyzed to compare cell migration and wound closure under different conditions. Introduction of different cytotoxic treatments into a wound healing assay can provide information as to whether a particular drug or compound of interest has the ability to affect cell migration. This type of analysis is important when assessing the ability of a particular cancer cell line to display invasive and metastatic behaviors. One of the challenges of this assay is to create the original wound in a way that is consistent across plates or treatments, facilitating comparisons across experimental groups. This is a particular challenge when using the wound healing assay in the context of an undergraduate biology class to expose students to a distinct form of mammalian cell culture and help them apply scientific knowledge and research skills. We found an easy way to overcome this obstacle by using ibidi plates. In this article, we provide a simple protocol to use ibidi plates and HeLa cells to set up wound healing assays. This laboratory exercise allows undergraduate students to utilize different skills developed through cell culture experience, such as growing, treating, and imaging mammalian cells.

摘要

伤口愈合实验是一种简单且成本低廉的方法,它能让研究人员通过实验模拟导致伤口愈合的细胞生长和迁移过程。在该实验中,在培养的哺乳动物细胞单层上制造一个伤口,并监测细胞迁移情况。在实验过程中定期拍摄显微照片。对这些显微镜图像进行分析,以比较不同条件下的细胞迁移和伤口闭合情况。在伤口愈合实验中引入不同的细胞毒性处理方法,可以提供有关特定药物或感兴趣的化合物是否有能力影响细胞迁移的信息。在评估特定癌细胞系表现出侵袭和转移行为的能力时,这种类型的分析很重要。该实验的挑战之一是以在不同平板或处理之间保持一致的方式制造原始伤口,以便于跨实验组进行比较。当在本科生物学课程中使用伤口愈合实验,让学生接触一种独特的哺乳动物细胞培养形式,并帮助他们应用科学知识和研究技能时,这是一个特别的挑战。我们发现了一种使用ibidi平板克服这一障碍的简单方法。在本文中,我们提供了一个使用ibidi平板和HeLa细胞进行伤口愈合实验的简单方案。这个实验室练习让本科生能够运用通过细胞培养经验培养的不同技能,如培养、处理和成像哺乳动物细胞。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/62f2/9429954/190876d60734/jmbe.00061-22-f001.jpg

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