Method for constructing in-vitro culture and damage model of pelodiscus sinensis embryonic period skirt fibroblasts

A fibroblast and in vitro culture technology, applied in cell dissociation methods, cell culture active agents, culture process, etc., can solve the problems of unreported effects of physiological effects, and achieve high efficiency and high cell survival rate and ratio , the effect of shortening the experiment time

Pending Publication Date: 2021-08-10
ZHEJIANG WANLI UNIV
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Problems solved by technology

At present, there are some reports about LPS on the immune-related genes, receptors, and related signaling pathways of soft-shelled turtles, but there are no reports on the effects of LPS on the specialized traits of soft-shelled turtles—skirt structure and physiological effects.

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  • Method for constructing in-vitro culture and damage model of pelodiscus sinensis embryonic period skirt fibroblasts
  • Method for constructing in-vitro culture and damage model of pelodiscus sinensis embryonic period skirt fibroblasts
  • Method for constructing in-vitro culture and damage model of pelodiscus sinensis embryonic period skirt fibroblasts

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Embodiment Construction

[0047] The present invention will be further described in detail below in conjunction with the accompanying drawings and embodiments.

[0048] In the present invention, unless specially prepared, the raw materials and equipment used can be purchased from the market or commonly used in the field. The methods in the following examples, unless otherwise specified, are conventional methods in the art.

[0049] Embodiment: the method for in vitro culture and injury model construction of soft-shelled turtle embryonic skirt fibroblasts comprises the following steps:

[0050] (1) Incubate fresh, normal-sized fertilized eggs of Chinese soft-shelled turtle in an incubator at 31°C and a relative humidity of 80% to 90%, and hatch them in an upright and naked state until the 17th to 20th stage of embryonic development. Take 6 After the surface of each fertilized egg was sterilized by immersing it in 75% alcohol for 15 seconds, the embryo was aseptically taken out with ophthalmic scissors ...

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Abstract

The invention discloses a method for constructing an in-vitro culture and damage model of pelodiscus sinensis embryonic period skirt fibroblasts. The method is characterized in that: the cells are derived from a key period of formation of the skirt character of a special nutritional tissue part of pelodiscus sinensis; and according to the characteristics of the hierarchical structure of the skirt organization, the cells have functional specificity of generation, secretion and deposition of different types of collagen. The method mainly comprises the following steps: primary culture, subculture, cell cryopreservation and skirt fibroblast damage model construction. The skirt fibroblasts are treated with lipopolysaccharide (LPS) with the concentration of 160 [mu]g / mL for 4 hours to obtain the skirt fibroblast damage model. The method has the advantages of being high in survival rate, good in operability, strong in characteristic feature, simple and practical. The pelodiscus sinensis embryonic period skirt cell system constructed in the invention can be used for researching the generation and physiological action of the pelodiscus sinensis skirt specialized character on the molecular cell level.

Description

technical field [0001] The present invention relates to the in vitro culture of the unique skirt tissue cells of Chinese soft-shelled turtle, in particular to a method for the in vitro culture of the embryonic skirt fibroblasts of the Chinese soft-shelled turtle and the construction of injury models. Background technique [0002] Chinese soft-shelled turtle, scientific name Pelodiscus sinensis, also known as soft-shelled turtle, turtle, water fish, etc., is a common special economic animal, belonging to the animal kingdom, chordate phylum, reptile class, turtle order, turtle family, soft-shelled turtle genus. China, Japan, northern Vietnam, eastern Russia, and South Korea all have wild Chinese soft-shelled turtles. In my country, it is more common in the Yangtze River Basin and South China. Chinese soft-shelled turtle is deeply loved by consumers at home and abroad because of its unique flavor, high edible value and medicinal value. [0003] Although turtles and turtles all...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N5/07
CPCC12N5/0601C12N2509/00C12N2509/10C12N2500/30C12N2501/90
Inventor 周贝宁李彩燕宋伟钱国英
Owner ZHEJIANG WANLI UNIV
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