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A method for primary culture of dorsal root ganglion satellite glial cells

A technology of dorsal root ganglion and glial cells, which is applied in the field of isolation and culture of dorsal root ganglion satellite glial cells, can solve the problems of failing to separate and culture satellite glial cells, and achieve the effect of large quantity and high purity

Active Publication Date: 2020-09-08
KUNMING MEDICAL UNIVERSITY
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  • Abstract
  • Description
  • Claims
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AI Technical Summary

Problems solved by technology

The above methods can only isolate and culture neurons from DRG in vitro, but fail to isolate and culture satellite glial cells

Method used

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  • A method for primary culture of dorsal root ganglion satellite glial cells
  • A method for primary culture of dorsal root ganglion satellite glial cells
  • A method for primary culture of dorsal root ganglion satellite glial cells

Examples

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Embodiment 1

[0031] 1 Materials and methods

[0032] 1.1 Experimental animals

[0033] SD neonatal rats born at 0-24 hours were purchased from the Department of Experimental Zoology of Kunming Medical University, and the invention was approved by the Medical Ethics Committee of Kunming Medical University.

[0034] 1.2 Take the equipment needed for DRG

[0035] Stereo microscope, ophthalmic microscopic straight tweezers, ophthalmic microscopic curved tweezers, small scissors, large scissors, flat pointed tweezers, large and small hairspring tweezers.

[0036] 1.3 Main reagents for DRG-SGC culture

[0037] DMEM / F12 (1:1) medium, B-27 supplement (50×), double antibody (penicillin and streptomycin solution), L-glutamine, BSA (30mg / mL), dexamethasone (25ug / mL), bovine insulin Insulin (5mg / mL) were purchased from biosharp company; T3 (10ug / mL), T4 (400ug / mL), NRG-β1 were purchased from abcam company, fetal bovine serum, and 0.25% trypsin were purchased from the United States Life Technology...

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Abstract

The invention discloses a primary culturing method for dorsal root ganglion satellite glial cells. The method comprises the operating steps of 1, killing a neonatal mouse or a neonatal rat after sterilization by breaking a head, taking out a spine after dissection, removing muscle on the spine, cutting off the spine, clearing blood vessels and spinal cord under a stereoscopic microscope and taking out DRG, and removing nerve fibers and envelopes on the DRG; 2, culturing the processed DRG by using DRG-SGCs culturing solution, inducing the large number of satellite glial cells to move out of the DRG, adding trypsin for digestion after culturing ends, then adding FBS for stopping digestion, slighting blowing and beating the solution by using a Pasteur pipet, and then transferring the solution into a centrifuge tube for centrifuging; and 3, discarding supernate after centrifuging, and then adding the culturing solution for continuous culturing, thus acquiring the satellite glial cells. The satellite glial cells cultured and purified according to the method provided by the invention can survive for a long time in vitro, is high impurity and stable in survival, and can be passaged for multiple generations and form a cell network.

Description

technical field [0001] The invention belongs to the technical field of separation and culture of dorsal root ganglion satellite glial cells, and in particular relates to a method for in vitro primary culture of dorsal root ganglion satellite glial cells. Background technique [0002] The nerve cells of the dorsal root ganglion (DRG) are the primary afferent neurons of the somatosensory, and their central processes enter the spinal cord to send out ascending fibers, and form synaptic connections with neurons in other parts, such as nerves with the thalamus form the spinothalamic tract, etc. These neurons and ascending fibers play an important role in the transmission of sensation from the periphery to the center. Satellite glial cells (satellite glial cels, SGCs) are an important glial cell in peripheral nerves, they are widely distributed in dorsal root ganglia (DRG) and trigeminal ganglia (TG) Inside. Satellite glial cells surround 1-2 neurons to form a glial cell sheath,...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N5/079
CPCC12N5/0622C12N2500/30C12N2500/32C12N2501/33C12N2501/998C12N2509/00
Inventor 李力燕郭建辉马微杨金伟王先斌李兴统王通通代云飞张泰李永进
Owner KUNMING MEDICAL UNIVERSITY
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