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Application of Tubastatin A in preparation of in-vitro activating agent for primordial follicles

A technology of primordial follicles and activators, applied in the direction of cell culture active agents, biochemical equipment and methods, germ cells, etc., can solve the problems of low success rate, hindering the application of primordial follicle activation in vitro, long operation time, etc.

Active Publication Date: 2021-04-23
CHINA AGRI UNIV
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  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, due to the low success rate of the in vitro activation of primordial follicles used in assisted reproduction and the long operation time (1-2 hours), patients often wait for the primordial follicles to be activated before undergoing transplantation. had to undergo a second operation) and other limiting factors hindered the clinical application of in vitro activation of primordial follicles

Method used

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  • Application of Tubastatin A in preparation of in-vitro activating agent for primordial follicles
  • Application of Tubastatin A in preparation of in-vitro activating agent for primordial follicles
  • Application of Tubastatin A in preparation of in-vitro activating agent for primordial follicles

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Experimental program
Comparison scheme
Effect test

Embodiment 1

[0029] Experimental Materials:

[0030] Newborn C57BL / 6J female mice, 5-day-old C57BL / 6J female mice, and 35-day-old C57BL / 6J female mice.

[0031] Experimental reagents:

[0032] Reagent A: ovarian basal medium, the specific formula is as follows, the preparation process must be aseptic.

[0033]

[0034] Reagent B: 4mM Tubastatin A (solvent is dimethyl sulfoxide)

[0035] Reagent C: sterile PBS

[0036] experimental method

[0037] 1. Add 1.2ml of Reagent A and 1.2μl of Reagent B to the 6-well plate in sequence, and then place it in a cell culture incubator to preheat at 37°C;

[0038] 2. Remove the ovarian cortex tissue of each day-old mouse and cut it into 1mm in Reagent C 3 After the small pieces were transferred to the preheated medium in step 1, they were placed in a cell culture incubator for 30 minutes to activate primordial follicles in vitro. Transfer the ovarian tissue to the preheated reagent A and continue to culture for 12 hours. Detect phosphorylated ...

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Abstract

The invention relates to the technical field of biology, in particular to an application of Tubastatin A in preparation of an in-vitro activating agent for primordial follicles. The invention provides the application of the small molecular drug Tubastatin A in preparation of a primordial follicle activated preparation. Experiments show that Tubastatin A promotes precursor granulosa cells to grow and differentiate into granulosa cells by improving mTOR total protein and phosphorylation level, and KITL secreted by Tubastatin A is increased at the same time. The ligand KITL is combined with a receptor on an oocyte membrane to activate a PI3K signal in an oocyte, up-regulate the phosphorylation level of AKT and promote the activation of primitive follicles, so that the number of primary follicles is increased.

Description

technical field [0001] The invention relates to the field of biotechnology, in particular to the application of Tubastatin A in the preparation of primordial follicle activators in vitro. Background technique [0002] Assisted reproductive technology is the most effective means of assisting pregnancy at this stage. However, a prerequisite for the current assisted reproductive technology for female patients is that there must be follicles in the ovaries that can respond to gonadotropins. However, there are not enough gonadotropin-responsive follicles in the ovaries of patients with premature ovarian failure, and special groups of patients who suffer from cancer after radiotherapy / chemotherapy and due to ovarian cryopreservation or ovary removal also face the problem of infertility. [0003] In order to solve the reproductive problems of this type of patients, new assisted reproductive technology and new means of utilization of follicle resources must be developed. In fact, ...

Claims

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Application Information

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IPC IPC(8): C12N5/075
CPCC12N5/0609C12N2501/405C12N2501/727
Inventor 夏国良王超张拓何美娜
Owner CHINA AGRI UNIV
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