Looking for breakthrough ideas for innovation challenges? Try Patsnap Eureka!

Method for culturing primary neurons in brain tissue cortical area and transfecting primary neurons with AAV (adeno-associated virus)

A neuron culture medium, virus transfection technology, applied in the direction of viruses, nervous system cells, viruses/phages, etc., can solve the problems of unstable expression, inability to integrate foreign genes, difficult cytotoxicity, etc., to achieve good growth state, The effect of fast transfection and simple method

Inactive Publication Date: 2018-07-20
ZHEJIANG UNIV
View PDF0 Cites 2 Cited by
  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, the cytotoxicity caused by a strong electric field is difficult to solve, and the foreign gene cannot be integrated into the host genome, resulting in unstable expression, which requires special electroporation equipment

Method used

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
View more

Image

Smart Image Click on the blue labels to locate them in the text.
Viewing Examples
Smart Image
  • Method for culturing primary neurons in brain tissue cortical area and transfecting primary neurons with AAV (adeno-associated virus)
  • Method for culturing primary neurons in brain tissue cortical area and transfecting primary neurons with AAV (adeno-associated virus)
  • Method for culturing primary neurons in brain tissue cortical area and transfecting primary neurons with AAV (adeno-associated virus)

Examples

Experimental program
Comparison scheme
Effect test

Embodiment Construction

[0043] The present invention will be further described below with reference to the drawings and embodiments.

[0044] The embodiment of the present invention is as follows:

[0045] According to the primary cortical neuron culture and transfection method of the present invention, the culture and transfection of primary neurons in the cortical area of ​​the tissue are specifically as follows:

[0046] 1. Petri dish coating

[0047] One day before neuron culture, spread four glass slides with a diameter of 12mm in a 35mm petri dish, add 2ml of 0.1mg / ml poly-L-lysine, and place in a refrigerator at 4 degrees for 12-15 hours. Take out the petri dish , Use a pipette to remove all poly-L-lysine in the ultra-clean table, and place it in the ultra-clean table to wait for the petri dish to dry naturally. After drying, add 2ml of the first culture medium and place it in the cell incubator for standby. The box is set to 37 degrees Celsius and a carbon dioxide concentration of 5%.

[0048] 2. Sep...

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
Login to View More

PUM

No PUM Login to View More

Abstract

The invention discloses a method for culturing primary neurons in a brain tissue cortical area and transfecting the primary neurons with an AAV (adeno-associated virus). The method comprises the following steps: brain of a fetal rat is extracted, and cortical tissue is separated and subjected to digestive treatment; tissue impurities are removed, and a cell suspension is prepared in a first culture solution; the cell suspension is inoculated in a petri dish and cultured in a cell incubator, and a primary neuron culture solution is obtained; the AAV is added to the primary neuron culture solution for neuron transfection; solution change is performed continuously after transfection. The cultured primary neurons are good in state, and the transfecting method is stable, efficient and rapid andcan be used for transfecting large gene fragments.

Description

Technical field [0001] The invention belongs to the field of biotechnology, and specifically relates to a method for culturing primary neurons in the cortical region of brain tissue and transfecting with Adeno-associated Virus (AAV). Background technique [0002] The cortical neurons cultured in vitro are similar in morphology and physiological characteristics to maternal tissues and can simulate the internal environment. Therefore, they have become an important experimental model for studying neurological disease mechanisms, drug efficacy, and functional genetic modification. Cell transfection refers to the technology of introducing foreign molecules such as DNA and RNA into eukaryotic cells. With the continuous development of molecular biology research, transfection has become a routine tool for studying and controlling eukaryotes. Therefore, the transfection of primary neurons is very important in the field of neurology. However, because neurons are highly differentiated cells...

Claims

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
Login to View More

Application Information

Patent Timeline
no application Login to View More
IPC IPC(8): C12N5/0793C12N15/864
CPCC12N5/0619C12N15/86C12N2750/14143C12N2800/107
Inventor 龚薇斯科黄理蒙
Owner ZHEJIANG UNIV
Features
  • Generate Ideas
  • Intellectual Property
  • Life Sciences
  • Materials
  • Tech Scout
Why Patsnap Eureka
  • Unparalleled Data Quality
  • Higher Quality Content
  • 60% Fewer Hallucinations
Social media
Patsnap Eureka Blog
Learn More

Browse by: Latest US Patents, China's latest patents, Technical Efficacy Thesaurus, Application Domain, Technology Topic, Popular Technical Reports.

© 2025 PatSnap. All rights reserved.Legal|Privacy policy|Modern Slavery Act Transparency Statement|Sitemap|About US| Contact US: help@patsnap.com