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Primary culture method in serum-free medium for fetal rat cortical neurons

A serum-free culture, serum-free culture medium technology, applied in the medical field, can solve the problems of serum culture method interference, influence observation, etc.

Inactive Publication Date: 2012-12-26
SUZHOU HEALTH COLLEGE
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Problems solved by technology

[0004] The purpose of the present invention is to provide a primary serum-free culture method for fetal mouse cortical neurons, which solves the problems of interference and observation in the serum culture method in the prior art

Method used

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Embodiment 1

[0029] Prepare materials:

[0030] DMEM high-glucose medium, trypsin, and HEPES were purchased from GIBCO;

[0031] Polylysine, insulin-transferrin-sodium selenite serum-free medium (ITS) were purchased from Sigma;

[0032] Neuron-specific enolase (NSE) and ready-to-use HIGH-SABC immunohistochemistry kit are products of Beijing Zhongshan Jinqiao Biotechnology Co., Ltd.

[0033] The primary serum-free culture method of a kind of fetal mouse cortical neuron of the present invention, concrete steps are as follows:

[0034] Step 1) Preparation of reagents:

[0035] DMEM high glucose medium 13.4g / L, NaHCO 3 2.2g / L, HEPES 3.75g / L, dissolved in triple distilled water in turn, distilled to 1000ml, sterilized by suction, and stored at -20°C;

[0036] Planting solution: by volume, DMEM high-glucose medium 90%, fetal bovine serum 10%;

[0037] Breeding solution: by volume, 99% of DMEM high-glucose medium, 1% of insulin-transferrin-sodium selenite serum-free me...

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Abstract

The invention discloses a primary culture method in a serum-free medium for fetal rat cortical neurons. According to the method, an insulin-transferrin-sodium selenite serum-free medium is used to culture the fetal rat cortical neurons, and neuron-specific enolase is used for dyeing to identify the fetal rat cortical neurons. According to the primary culture method in a serum-free medium for the fetal rat cortical neurons, dissociated culture model of Wistar fetal rat cortical in vivo is established by using a serum-free culture method, and morphologic observation of the cultured neurons is carried out, establishing a foundation for relative researches in future.

Description

technical field [0001] The invention relates to the medical field, in particular to a method for primary serum-free culture of fetal rat cortical neurons. [0002] Background technique [0003] The complete medium for culturing cells consists of basal medium (such as MEM) and additives (such as serum or certain hormones and growth factors for serum-free culture). Most cells do not survive for a long time when only basal medium is used for cell culture. In order to promote their survival and growth, additional factors must be added, such as hormones, transporters and some trace elements. In general, serum is chosen to provide these factors. However, the composition of serum is complex and there are many unknown factors, which sometimes interfere with experimental results and cover up the potential physiological functions of cultured cells. In addition, under physiological conditions, nervous tissue is not directly exposed to most components of plasma due to the presenc...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N5/073C12N5/0793
Inventor 叶蓓
Owner SUZHOU HEALTH COLLEGE
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