Using a Primary Cell Culture Model to Study the Neural Extracellular Matrix
Using a Primary Cell Culture Model to Study the Neural Extracellular Matrix
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使用原代细胞培养模型研究神经细胞外基质
DOI：
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发表时间：
2020
期刊：
影响因子：
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通讯作者：
K. A. Giamanco
中科院分区：
文献类型：
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作者：
K. A. Giamanco
细胞外基质(ECM)为细胞提供结构支持，但在调节细胞黏附、细胞信号转导和分化方面也起着关键作用。虽然这个话题是在课堂上讨论的，但在实验环节中并没有深入研究。这一课是为了让学生通过使用大脑中的细胞外基质作为模型来理解细胞外基质是如何分子结构的。更具体地说，学生学习神经ECM中的一种特殊结构，称为神经周围神经网络(PNN)。在这项练习中，学生们研究大脑中哪种细胞类型有助于产生这种结构，以及活动的变化是否会影响PNN的发育。在这节课中，学生们分离和培养来自胚胎小鼠大脑的单个细胞；然后他们进行标准免疫荧光染色程序的各个方面。最后，学生们将使用荧光显微镜对培养物进行成像。所有上述的基本技术都被细胞生物学家广泛使用。完成后，学生们交上一份正式的实验报告，这份报告是根据主要文献进行背景分析的。学生们非常喜欢这个练习，他们发现它具有挑战性，但很有意义，发人深省。引文：Giamanco Ka。2020年。采用原代细胞培养模型研究神经细胞外基质。课程来源。Https://doi.org/10.24918/cs.2020.9编辑：斯科特·格勒，奥古斯塔纳学院收到日期：2019年7月9日；接受日期：2020年1月30日；发布日期：2020年5月28日。版权所有：©2020 Giamanco。这是一篇开放获取的文章，根据知识共享署名-非商业性-类似共享4.0国际许可证的条款分发，该许可证允许在任何媒体上不受限制地非商业性使用、分发和复制，前提是原始作者和来源必须注明身份。提交人确认，她拥有或已获得使用文本、图表、表格、插图、摘要、摘要和补充材料的书面许可。利益冲突和资金声明：作者没有与本作品相关的财务、个人或职业利益冲突。配套材料：S1：学习神经细胞外基质-实验1作业；S2：研究神经细胞外基质-实验3作业；S3：研究神经细胞外基质-解剖说明；S4：研究神经细胞外基质-实验1讲义；S5：研究神经细胞外基质-实验2实验前讲义；S6：研究神经细胞外基质-实验2讲义；S7：研究神经细胞外基质-实验3实验前讲义；S8：研究神经细胞外基质-实验3讲义；S9：研究神经细胞外基质-实验4实验前讲义；S10：研究神经细胞外基质-实验讲义4；S11：研究神经细胞外基质-教师笔记；和S12：研究神经细胞外基质-实验报告和题目信息。*通信地址：美国康涅狄格州丹伯里205号科学大楼白街181号邮编06810美国邮编：giamancok@wcsu.edu
The extracellular matrix (ECM) provides structural support to cells, but also has key roles in mediating cell adhesion, cell signaling, and differentiation. While this topic is discussed in the lecture setting, it is not heavily studied in laboratory sessions. This lesson was created so that students can understand how the ECM is molecularly structured by using the ECM in the brain as a model. More specifically, the students study a particular structure in the neural ECM called the perineuronal net (PNN). In this exercise, students examine which cell type in the brain helps produce this structure and whether changes in activity affect PNN development. In this lesson students isolate and plate single cells derived from embryonic mouse brains; they then perform aspects of a standard immunofluorescence staining protocol. Lastly, the students will image the cultures using a fluorescence microscope. All of the aforementioned fundamental techniques are widely used by cell biologists. After completion, students hand in a formal lab report that is contextualized by primary literature. Students thoroughly enjoyed this exercise, and they found it challenging but rewarding and thought provoking. Citation: Giamanco KA. 2020. Using a primary cell culture model to study the neural extracellular matrix. CourseSource. https://doi.org/10.24918/cs.2020.9 Editor: Scott Gehler, Augustana College Received: 7/9/2019; Accepted: 1/30/2020; Published: 5/28/2020 Copyright: © 2020 Giamanco. This is an open-access article distributed under the terms of the Creative Commons Attribution-NonCommercial-ShareAlike 4.0 International License, which permits unrestricted non-commercial use, distribution, and reproduction in any medium, provided the original author and source are credited. Author affirms that she either owns the copyright to or has received written permission to use the text, figures, tables, artwork, abstract, summaries and supplementary materials. Conflict of Interest and Funding Statement: The author does not have a financial, personal, or professional conflict of interest related to this work. Supporting Materials: S1: Studying the Neural Extracellular Matrix – Lab Session 1 Assignment; S2: Studying the Neural Extracellular Matrix – Lab Session 3 Assignment; S3: Studying the Neural Extracellular Matrix – Dissection Instructions; S4: Studying the Neural Extracellular Matrix – Lab Session 1 Handout; S5: Studying the Neural Extracellular Matrix – Lab Session 2 Pre-Lab Lecture; S6: Studying the Neural Extracellular Matrix – Lab Session 2 Handout; S7: Studying the Neural Extracellular Matrix – Lab Session 3 Pre-Lab Lecture; S8: Studying the Neural Extracellular Matrix – Lab Session 3 Handout; S9: Studying the Neural Extracellular Matrix – Lab Session 4 Pre-Lab Lecture; S10: Studying the Neural Extracellular Matrix – Lab Session 4 Handout; S11: Studying the Neural Extracellular Matrix – Instructor Notes; and S12: Studying the Neural Extracellular Matrix – Lab Report and Rubric Information. *Correspondence to: 181 White Street, Science Building 205, Danbury, CT 06810 USA email: giamancok@wcsu.edu CourseSource | www.coursesource.org 2020 | Volume 07 1 Lesson
DOI：
10.3791/3965
发表时间：
2012-05-24
期刊：
Journal of visualized experiments : JoVE
影响因子：
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作者：
Pacifici M;Peruzzi F
通讯作者：
Peruzzi F