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Separation and culture method of human retina Muller cells

A culture method and retinal technology, applied in the field of cell culture in modern biotechnology, can solve the problems of little research on retinal Muller cell culture methods, difficult access to eyeball tissue, and restrictions on research work, so as to achieve simple and easy-to-master technology, cell The effect of good growth status and reliable cell resources

Inactive Publication Date: 2019-10-29
JIANGYIN CHI SCI
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  • Abstract
  • Description
  • Claims
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Problems solved by technology

[0003] At present, there are very few human retinal Muller cells available at home and abroad, and there are few studies on normal retinal Muller cell culture methods in vitro, and most of the methods are unstable. In addition, the eyeball tissue is not easy to obtain, which seriously restricts the follow-up research work

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  • Separation and culture method of human retina Muller cells
  • Separation and culture method of human retina Muller cells
  • Separation and culture method of human retina Muller cells

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Embodiment Construction

[0016] The terms used in the present invention, unless otherwise specified, generally have the meanings commonly understood by those skilled in the art.

[0017] The present invention will be described in detail below in conjunction with the accompanying drawings and examples, and the protection content of the present invention is not limited to the following examples.

[0018] In the following examples, various procedures and methods not described in detail are conventional methods well known in the art.

[0019] The experimental instruments and reagents used in this experiment are as follows: a set of surgical instruments, an inverted microscope (XDS-1A, Shanghai), a fluorescence microscope (Leica, the United States), a constant temperature incubator (Heraeus, the United States), a low-temperature centrifuge (TD24B-WS , Shanghai), ultra-low temperature refrigerator (Zhongke Meiling), pipette gun (Eppendorf, USA), electronic analytical balance (Sartorius, USA), ultra-clean be...

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Abstract

The invention provides a separation and culture method of primary human retina Muller cells. The separation and culture method comprises the following steps: (1) a normal eyeball surgically excised from a corneal donor is taken out aseptically; (2) under a dissecting microscope, retina tissue is detached aseptically; (3) mixed collagenase is added for digestion; (4) digestion is terminated, filtering, centrifuging and washing are conducted, re-suspending is conducted through a complete medium, and the complete medium is inoculated into a culture bottle; (5) the culture bottle is placed into a5% CO2 incubator at 37 DEG C to be cultured; (6) cell passage and purification are conducted; and (7) the cell morphology is observed and identified. According to the separation and culture method ofthe human retina Muller cells, operation is easy, stability and high efficiency are achieved, and a good experimental material is provided for further studying a retinopathy damage generation mechanism.

Description

technical field [0001] The invention belongs to the technical field of cell culture of modern biotechnology, in particular to a method for separating and culturing human retinal Muller cells. Background technique [0002] Muller cells were first described by German Müller (1851) and named after him. Because of its elongated shape, it is also called Muller fiber. Its cell body emits slender protrusions to the inside and outside, occupying the entire thickness of the retina from the inner limiting membrane to the outer limiting membrane, surrounding most of the cones, rods, bipolar cells and ganglion cells in the retina, and specialized foot plates Attached to the retinal capillary wall to participate in the formation of the blood - retinal barrier. Muller cells are the main glial cells in the vertebrate retina. It runs through the entire retina and interacts with retinal nerve cells and retinal blood vessels in multiple functions. The pathogenesis of diabetic retinopathy ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N5/079
CPCC12N5/0621C12N2509/00
Inventor 不公告发明人
Owner JIANGYIN CHI SCI
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