Method for adjusting and controlling ovarian granular cells based on circ-Atp5BP

A technology of granulosa cells and ovaries, applied in the field of ovarian granulosa cell regulation based on circ-Atp5BP, can solve the problems of tissue specificity and spatiotemporal specificity of circRNA, and achieve the effect of regulating proliferation.

Inactive Publication Date: 2021-06-22
SICHUAN AGRI UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

[0006] At present, a large number of studies have shown that compared with lncRNA, circular RNA has better stability and conservation, and it is not difficult to find that circRNA has tissue specificity and spatiotemporal specificity through the calculation of data such as sequencing.

Method used

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  • Method for adjusting and controlling ovarian granular cells based on circ-Atp5BP
  • Method for adjusting and controlling ovarian granular cells based on circ-Atp5BP
  • Method for adjusting and controlling ovarian granular cells based on circ-Atp5BP

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Embodiment 1

[0034] 1. Experimental plan

[0035] 1. Isolation and culture of porcine ovary granulosa cells

[0036] After the sows were slaughtered, the ovaries were taken out, washed in PBS, stored in saline at 38 degrees Celsius, and brought back to the laboratory within 1 hour to separate the ovarian granulosa cells.

[0037] Put the ovary in the preheated 37-degree normal saline and clean it, use a scalpel to cut the surface of the ovary evenly and lightly at intervals of 1mm, collect the follicle fluid, put the absorbed follicle fluid into a 15ml centrifuge tube, 1000rpm Centrifuge for 8 minutes, discard the supernatant, gently pipette with preheated PBS to evenly wash and centrifuge twice.

[0038] The prepared ovarian granulosa cell culture medium is 20% FBS plus DMEM and 1% third antibody (penicillin, streptomycin and amphotericin)

[0039] 2. Cell inoculation, subculture, transfection, and sample collection

[0040] When the ovarian granulosa cells proliferate to 85%, discard ...

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Abstract

The invention provides a method for adjusting and controlling ovarian granulosa cells based on circ-Atp5BP. The method comprises the following steps: (1) carrying out inoculation and subculture on nest granulosa cells; and (2) when the cell density is cultured to be 35 to 45 percent, transfecting the circ-Atp5BP gene or the pik3 gene into the nest granular cells. The application of circ-Atp5BP and pik3 in porcine ovarian granular cell proliferation adjustment and the targeting adjustment relation between circ-Atp5BP and pi3k are proved for the first time. The proliferation of the ovarian granular cells can be remarkably adjusted and controlled.

Description

technical field [0001] The invention belongs to the technical field of cell engineering and genetic engineering, and specifically relates to a method for regulating ovarian granulosa cells based on circ-Atp5BP. Background technique [0002] The ovary is an important organ that determines the fertility of female animals, and one of its main functions is follicle development and ovulation. Abnormal ovarian function can lead to reduced reproduction, including premature ovarian failure (Premature ovarian failure, POF), polycystic ovarian syndrome (polycystic ovarian syndrome, PCOS) and ovarian cancer. Among them, the main causes of premature ovarian failure are: first, not enough primordial follicles are formed during the embryonic period (congenital deficiency); second, the activation and inhibition of primordial follicles are affected, and some studies have shown that the acceleration of follicular atresia can also cause premature ovarian failure. [0003] During the developm...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N5/10C12N15/113
CPCC12N5/0682C12N15/113C12N2310/532C12N2510/00
Inventor 张锫文朱砺张顺华沈林園牛丽莉赵叶陈蕾吴霜郭鑫宇李欣荣肖茜林许韬
Owner SICHUAN AGRI UNIV
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