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Preparation method of rat corneal limbus stem cell deficiency model

A technology of limbal stem cell deficiency, which is applied in animal husbandry and other fields, can solve the problems that the degree of damage is difficult to control, it is difficult to ensure the uniformity of the pathological model of limbal stem cell deficiency, etc., and achieve the effect of reliable induction and precise placement time

Inactive Publication Date: 2021-06-11
XIEHE HOSPITAL ATTACHED TO TONGJI MEDICAL COLLEGE HUAZHONG SCI & TECH UNIV
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Problems solved by technology

[0003] The rat limbal stem cell deficiency model is the most common animal model to study the pathophysiological changes of the cornea and conjunctiva caused by the lack of corneal limbal epithelial stem cells caused by various types of inflammation and injury on the ocular surface and the response to various therapeutic measures. The traditional method for preparing the model is Putting the filter paper soaked in sodium hydroxide on the surface of the limbus caused damage to the limbal epithelial stem cells. However, since the surface of the eyeball of the rat is spherical and the filter paper is flat, it is difficult to infiltrate each limbus of the rat with the traditional filter paper. The location and degree of damage are not easy to control, and it is difficult to ensure the unity of the pathological model of limbal stem cell deficiency

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  • Preparation method of rat corneal limbus stem cell deficiency model
  • Preparation method of rat corneal limbus stem cell deficiency model
  • Preparation method of rat corneal limbus stem cell deficiency model

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Embodiment 1

[0025] Embodiment 1: A method for preparing a rat limbal stem cell deficiency model, comprising the following steps: S1: using pentobarbital sodium intraperitoneal injection to anesthetize experimental rats, and then dripping 4 mg / mL procaine on the rats for the experiment eye for local anesthesia; the diameter of the experimental eye of the rat in S1 is 5 mm; S2: the conical filter paper can be used to infiltrate all parts of the rat corneal limbus at the same time, and the time for placing the filter paper can be more accurately controlled; the conical filter paper in S2 Prepared by the following steps: Step 1: First cut the experimental qualitative filter paper into a semicircle with a diameter of 14 mm; Step 2: Fold the semicircular filter paper in half to form a cone with a bottom radius of 3.5 mm, and use double-sided tape Bond the mating surfaces to obtain a conical filter paper; S3: Soak the conical filter paper prepared in S2 in sodium hydroxide solution, clamp the top...

Embodiment 2

[0029] Embodiment 2: A method for preparing a rat limbal stem cell deficiency model, comprising the following steps: S1: using pentobarbital sodium intraperitoneal injection to anesthetize experimental rats, and then dripping 4 mg / mL procaine on the rats for the experiment The eye was used for local anesthesia; the diameter of the experimental eye of the rat in S1 was 5 mm; in S1, the experimental rat was anesthetized by intraperitoneal injection of 3% pentobarbital sodium 30 mg / kg, and procaine 4 mg / mL; S2: using conical The filter paper can simultaneously infiltrate various parts of the limbus of rats, and can more accurately control the placement time of the filter paper; the conical filter paper in S2 is prepared by the following steps: Step 1: First cut the experimental qualitative filter paper into a semicircle with a diameter of 14mm ;Step 2: Fold both sides of the semicircular filter paper in half to form a conical shape with a bottom radius of 3.5 mm, and use double-si...

Embodiment 3

[0033] Embodiment 3: A method for preparing a rat limbal stem cell deficiency model, comprising the following steps: S1: using pentobarbital sodium intraperitoneal injection to anesthetize experimental rats, and then dripping 4 mg / mL procaine on the rats for the experiment The eye was used for local anesthesia; the diameter of the experimental eye of the rat in S1 was 5.6 mm; in S1, the experimental rat was anesthetized by intraperitoneal injection of 3% pentobarbital sodium 30 mg / kg, and the procaine was 4 mg / mL; S2: using cone Shaped filter paper, which can infiltrate all parts of the rat corneal limbus at the same time, can control the time of filter paper placement more precisely; the conical filter paper in S2 is prepared by the following steps: Step 1: First cut the experimental qualitative filter paper into a semicircle with a diameter of 14mm shape; Step 2: Fold the semicircular filter paper in half to form a conical shape with a bottom radius of 3.5 mm, and use double-...

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Abstract

The invention discloses a preparation method of a rat corneal limbus stem cell deficiency model in the field of corneal epithelium. The method comprises the following steps of S1, anesthetizing an experimental rat by adopting pentobarbital sodium intraperitoneal injection, and dripping 4 mg / mL of procaine into the experimental eye of the rat for local anesthesia; S2, by utilizing conical filter paper, infiltrating all parts of the corneal limbus of the rat at the same time, and controlling the placement time of the filter paper more accurately; and S3, soaking the conical filter paper prepared in the step S2 in a sodium hydroxide solution, clamping the top of the conical filter paper through tweezers, and slightly sucking redundant liquid on the conical filter paper through gauze. By utilizing the conical filter paper, all the parts of the corneal limbus of the rat can be infiltrated at the same time, and the placement time of the filter paper can be more accurately controlled; compared with a traditional plane filter paper method, the preparation method has the advantages that the rat corneal limbus stem cell deficiency can be more stably and reliably induced, a good animal model for simulating a clinical patient is provided for experimental research, and a foundation is laid for further therapeutic research.

Description

technical field [0001] The invention relates to the field of corneal epithelium, in particular to a method for preparing a rat corneal limbal stem cell deficiency model. Background technique [0002] Limbal stem cell deficiency is a general term for a group of diseases characterized by severe limbal injury leading to a reduction in the number of available stem cells, which is insufficient for a continuous supply of corneal epithelial regeneration. Limbal stem cell deficiency will lead to corneal clouding, conjunctival and Vascularization, which destroys the transparency of the cornea, affects vision and eventually leads to vision loss. [0003] The rat limbal stem cell deficiency model is the most common animal model to study the pathophysiological changes of the cornea and conjunctiva caused by the lack of corneal limbal epithelial stem cells caused by various types of inflammation and injury on the ocular surface and the response to various therapeutic measures. The tradit...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A01K67/027
CPCA01K67/027A01K2207/20A01K2227/105A01K2267/03
Inventor 肖宇婷赵心悦胡芷馨柴晨汪晨扬
Owner XIEHE HOSPITAL ATTACHED TO TONGJI MEDICAL COLLEGE HUAZHONG SCI & TECH UNIV
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