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Application of cdc42 signaling pathway in activation of primordial follicle growth and development

A primordial follicle, growth and development technology, applied in the direction of microorganisms, germ cells, cell culture active agents, etc., can solve the problems of safety that need to be further evaluated, and achieve the effect of promoting development

Active Publication Date: 2021-03-16
CHINA AGRI UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

At the same time, currently commonly used activators of PI3K signaling pathway or mTORC1 signaling pathway are generally chemically synthesized molecules, and since activators of PI3K signaling pathway or mTORC1 signaling pathway are directly related to cancer, their safety needs to be further evaluated

Method used

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  • Application of cdc42 signaling pathway in activation of primordial follicle growth and development
  • Application of cdc42 signaling pathway in activation of primordial follicle growth and development
  • Application of cdc42 signaling pathway in activation of primordial follicle growth and development

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Experimental program
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Effect test

Embodiment 1

[0041] Embodiment 1: The reagent (in vitro) for activating the growth and development of primordial follicles according to the present invention

[0042] Solution I: basically activated cell culture medium;

[0043] Solution Ⅱ: DMEM / F12 culture medium is the mother solution, containing ITS;

[0044] CDC42 pathway activator: EGF;

[0045] The reagent of the present invention can pack solution I, solution II and the CDC42 pathway activator separately, and pack them together in the same packaging box, wherein the solution I and the CDC42 pathway activator are mixed to become a reagent for activating primordial follicles, and solution II is activated in vitro Used for cleaning primordial follicles;

[0046] The preparation method of the primordial follicle activation reagent in vitro, solution I: the preparation method of the basic activated cell culture medium is as follows: DMEM / F12 is the mother solution, containing 100 times of ITS, 10 Units / ml streptomycin and 10 g / ml penic...

Embodiment 2

[0052] Embodiment 2: The agent of the present invention for activating the growth and development of primordial follicles (in vivo)

[0053] CDC42 pathway activator: EGF;

[0054] Water-soluble gel: matrix hydrosol matrigel or cross-linked hyaluronic acid gel for uterine cavity, this embodiment uses matrix hydrosol matrigel.

[0055] The reagent of the present invention can pack the CDC42 pathway activator and the water-soluble gel separately, and pack them together in the same packaging box. When used, the concentration of the CDC42 pathway activator (EGF) is 50-200ng / ml, and is dissolved in the hydrosol , use 100ng / ml for specific application.

[0056] The method of using the reagent is as follows:

[0057] 1) Under sterile conditions, fully mix the reagent with the absorbable cross-linked hyaluronic acid gel for uterine cavity;

[0058] 2) During the operation, the gel mixed with the reagent is applied to the surface of the ovary by intracorporeal injection;

[0059] 3)...

Embodiment 3

[0060] Example 3: Effect of CDC42 Signaling Pathway on Primordial Follicle Activation and Growth and Development

[0061] 1. CDC42 inhibition experiment

[0062] Experimental animals: The animals used in the experiment were newborn 6-day-old C57 mice.

[0063] Inhibitors: CDC42 pathway inhibitor ML141 was purchased from Sigma, and ZCL278 was purchased from Selleck.

[0064] Two CDC42 inhibitors, ML141 and ZCL278, were used to treat neonatal mouse ovaries cultured in vitro for 3 days, and then tissue sections and germ cell-specific immunofluorescence staining were used to detect the development of primordial follicles in the ovary, and immunoblotting was used to detect the post-treatment Changes in CDC42 activity in the ovary, and follicle activation was counted by follicle counting. see results figure 1 .

[0065] Depend on figure 1 It can be seen that the inhibition of CDC42 activity leads to the blockage of primordial follicle activation. The results in A show that the...

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Abstract

The invention relates to the technical field of biology and discloses application of a CDC42 signal channel in activation of growth and development of primordial follicles. A CDC42 channel inhibitor and a CDC42 over-expression vector are adopted to confirm that the primordial follicles can be activated by quickly activating CDC42, and the development of the primordial follicles can be promoted, thereby providing a bran-new signal channel for activation and development of the primordial follicles. A CDC42 activator and the CDC42 over-expression vector can be applied to reagents for activating growth and development of the primordial follicles and / or applied to in vitro activation of growth of the primordial follicles for non-therapeutic purposes.

Description

technical field [0001] The invention relates to the field of biotechnology, in particular to the application of CDC42 signaling pathway in activating the growth and development of primordial follicles. Background technique [0002] Mammalian ovary is a reproductive organ necessary for the reproduction of female animals including human females. The ovary has two main functions: supporting the development and maturation of oocytes and producing female hormones. From birth to development, childbearing age, and finally to perimenopause and old age, the ovary undergoes a series of changes, which directly affect reproductive function, sexual characteristics and psychology. [0003] In the process of female germ cell development, the primordial follicle is the most basic functional unit of the ovary. The primordial follicle consists of a single dormant oocyte surrounded by a layer of flattened pregranulosa cells. The number of primordial follicles, that is, the primordial follicl...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N5/075
CPCC12N5/0609C12N2501/00C12N2501/11
Inventor 张华张佳伟颜昊夏国良
Owner CHINA AGRI UNIV
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