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JoVE Journal
Immunology and Infection
原生动物寄生虫感染原发性星细胞和微胶质的伴随分离
原生动物寄生虫感染原发性星细胞和微胶质的伴随分离
JoVE Journal
Immunology and Infection
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JoVE Journal Immunology and Infection
Concomitant Isolation of Primary Astrocytes and Microglia for Protozoa Parasite Infection

原生动物寄生虫感染原发性星细胞和微胶质的伴随分离

Full Text
8,202 Views
09:34 min
March 18, 2020

DOI: 10.3791/60680-v

Aline de Oliveira Lima Pacheco*1, Marcelo Pires Amaral*1, Ingrid Sancho de Farias1, Luiza Zainotti Miguel Fahur Bottino1, Karina Ramalho Bortoluci2

1Centro de Terapia Celular e Molecular (CTCMol),Universidade Federal de São Paulo (UNIFESP), 2Departamento de Ciências Biológicas, Centro de Terapia Celular e Molecular (CTCMol),Universidade Federal de São Paulo (UNIFESP)

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Overview

This protocol outlines a method for the extraction and dissociation of murine astrocyte and microglia cells from the central nervous system. It emphasizes the simultaneous isolation of both cell types to enhance experimental efficiency and comparative analysis in immunological studies.

Key Study Components

Area of Science

  • Neuroscience
  • Cell Biology
  • Immunology

Background

  • Astrocytes and microglia are the most abundant glial cells in the central nervous system.
  • Understanding their roles is crucial in studying diseases like Alzheimer��s and Parkinson��s.
  • Simultaneous extraction of both cell types can provide insights into their interactions during infections.
  • Previous methods often isolated these cells separately, which can be less efficient.

Purpose of Study

  • To develop a protocol for the simultaneous extraction of astrocytes and microglia.
  • To optimize the use of experimental animals in research.
  • To facilitate the study of the roles of these cells in various diseases and infections.

Methods Used

  • Extraction of glial cells from postnatal mice.
  • Isolation of astrocytes and microglia in parallel.
  • Careful handling during brain removal and tissue washes.
  • Time investment of approximately four hours for extraction.

Main Results

  • Successful isolation of both astrocytes and microglia from the same extraction.
  • Improved understanding of the comparative roles of these cells in immunological contexts.
  • Potential applications in studying infections by protozoa and viruses.
  • Enhanced efficiency in the use of experimental animals.

Conclusions

  • The protocol provides a reliable method for extracting glial cells.
  • It supports comparative studies of astrocytes and microglia.
  • Future research can benefit from this method in various disease contexts.

Frequently Asked Questions

What are astrocytes and microglia?
Astrocytes and microglia are the two most abundant glial cell types in the central nervous system, playing crucial roles in brain health and disease.
Why is simultaneous extraction important?
Simultaneous extraction allows for a more efficient use of experimental animals and provides insights into the interactions between these cell types.
How long does the extraction process take?
The extraction process typically takes about four hours, depending on the number of animals used.
What precautions should be taken during the protocol?
Care should be taken during brain removal and tissue washes to ensure the integrity of the cells.
In what contexts can this protocol be applied?
This protocol can be applied in studies related to infections, inflammation, and neurodegenerative diseases.

该协议的总体目标是指导如何从中枢神经系统中提取、维持和分离鼠星细胞和微胶质细胞,然后感染原生动物寄生虫。

该协议描述了一种方法,专注于从产后小鼠中同时提取两个最丰富的胶质细胞群,星细胞和微胶质。我们的方法不同于之前描述的星细胞和微胶质分离协议,因为我们专注于在同一条件下获得和隔离同一提取中的两种细胞类型,从而优化了实验动物的使用,并改进了这些细胞在免疫学背景下的相对作用的研究。该协议可用于更好地研究微胶质和星细胞作用,在许多情况下和疾病在中枢神经系统,如阿尔茨海默氏症,帕金森,和感染,例如原生动物寄生虫或病毒。

从同一萃取中获取和隔离两种细胞类型是评估每个细胞子集免疫学背景(如感染或炎症)的比较作用的可取的。人们应该小心,特别是在脑切除和神经组织洗涤,但我相信,有了这个协议和视频拍摄,人们不会挣扎。该协议可以很长,这取决于提取皮质的动物的数量,所以准备花费约四个小时或更多时间。

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