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Chronic glaucoma animal model establishing method and application thereof

An animal model and method of establishing a technology, applied in animal cells, vertebrate cells, pharmaceutical formulations, etc., can solve problems such as short time, and achieve low cost and good repeatability.

Inactive Publication Date: 2018-06-29
XIAMEN EYE CENTER OF XIAMEN UNIVERSITY CO LTD
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Problems solved by technology

Although these methods can increase the intraocular pressure of experimental animals in the early stage of modeling, the establishment of these chronic glaucoma animal models mostly requires high technical conditions, and even the time for maintaining high intraocular pressure in animals is relatively short

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  • Chronic glaucoma animal model establishing method and application thereof
  • Chronic glaucoma animal model establishing method and application thereof
  • Chronic glaucoma animal model establishing method and application thereof

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Embodiment Construction

[0036] The chronic glaucoma animal model establishment method disclosed by the present invention comprises the following steps:

[0037] (1) Take the conjunctival tissue of GFP-SD rats, wash it twice with sterilized PBS; put the conjunctival tissue into Dispase II or type IV collagenase digestion solution, and place it in a 37°C cell culture incubator Digest for 2 hours or overnight at 4°C, separate and discard the conjunctival epithelial layer, and obtain conjunctival stromal tissue pieces;

[0038] (2) The above-mentioned conjunctival stromal tissue pieces were digested in trypsin for 10 min, then digested with cell culture medium (DMEM medium containing 10% FBS, 1% streptomycin, and 1% penicillin), and blown into single cells Suspension: the above suspension was passed through a cell sieve with a pore size of 70um, centrifuged at 1500rpm in a centrifuge at 4°C for 5min, and the supernatant was discarded to obtain a conjunctival stromal cell pellet;

[0039] (3) Add an appr...

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Abstract

The invention discloses a chronic glaucoma animal model establishing method. The method comprises the following steps: (1) collecting conjunctiva tissues of GFP-SD rats, sterilizing, digesting, separating a conjunctiva epithelial layer, and obtaining a conjunctiva matrix tissue block; (2) digesting the conjunctiva matrix tissue block in pancreatin, terminating the digestion by using a cell culturesolution, blowing and beating to form a single-cell suspension, filtering by virtue of a cell sieve, centrifuging, and removing supernatant, thus obtaining conjunctiva matrix cell precipitates; (3) adding an appropriate amount of cell culture solution, blowing and beating to form single cells, counting, performing the in-vitro culture, and obtaining a remarkable amount of conjunctiva matrix cells; (4) digesting the conjunctiva matrix cells by using pancreatin, processing by utilizing the method of the step (2), obtaining the conjunctiva matrix cell precipitate, adding an appropriate amount ofcell culture solution, and obtaining cell suspension; and (5) injecting 10 micro liter of cell suspension by utilizing a micro injector into the anterior chamber of the right eye of a rat, and sufficiently and uniformly mixing. A glaucoma animal model obtained by using the method has the advantages of long-term stability, simplicity, ease, good repetition, and low cost.

Description

technical field [0001] The invention relates to a method for establishing a chronic glaucoma animal model and its application. Background technique [0002] Glaucoma is a group of diseases characterized by optic atrophy, visual field defect and vision loss. Pathological intraocular pressure increase and optic nerve insufficiency are the primary risk factors for its onset. The tolerance of optic nerve to pressure damage is also related to the occurrence of glaucoma. related to development. As of 2010, there are about 30 million glaucoma patients worldwide, and the prevalence rate of people over the age of 40 is 3.5% ([1] Glaucoma (2017). The Lancet. 2017; 390: 2183-2193). Glaucoma is one of the three leading causes of blindness in the world, seriously affecting people's quality of life. [0003] Glaucoma animal models play an important role in better understanding the damage mechanism and treatment measures of glaucomatous optic neuropathy. At present, the animal models of...

Claims

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Application Information

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IPC IPC(8): A61K35/30C12N5/079A61K49/00A01K67/02
CPCA61K35/30A01K67/02A01K2227/105A01K2267/03A61K49/0008C12N5/0621
Inventor 吴仁毅黄昌泉汪耀吕洁璇李晓红
Owner XIAMEN EYE CENTER OF XIAMEN UNIVERSITY CO LTD
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