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Application of n-acetylcysteine ​​in the preparation of porcine placenta trophoblast cell in vitro culture medium

A technology of acetylcysteine ​​and trophoblast cells is applied in the field of livestock breeding to achieve the effect of promoting proliferation

Active Publication Date: 2021-08-17
BEIJING ACADEMY OF AGRICULTURE & FORESTRY SCIENCES
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0005] NAC has broad application prospects in animal husbandry production, but so far, there is no report on the effect of NAC on porcine placental trophoblast cells

Method used

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  • Application of n-acetylcysteine ​​in the preparation of porcine placenta trophoblast cell in vitro culture medium
  • Application of n-acetylcysteine ​​in the preparation of porcine placenta trophoblast cell in vitro culture medium

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0024] The effect of embodiment 1N-acetylcysteine ​​on pig placental trophoblast cell proliferation

[0025] 1. Cell culture

[0026] Pig placental trophoblast cells pTr (porcine trophectoderm cells) were provided by Animal Nutrition Laboratory of China Agricultural University. After thawing in a water bath at 37°C, suck out the cells and add 2 mL of complete culture solution (100 mL of complete culture solution contains 88 mL of DMEM:F12, 10 mL of fetal bovine serum, 1 mL of ITS and 1 mL of penicillin / streptomycin double antibody solution) Mix well, centrifuge at 1000 rpm for 4 minutes, discard the supernatant, resuspend with 2 mL of complete culture medium and place in a 75T cell culture flask in 5% CO 2 Cultured under 95% air conditions, cultured for 48 hours or when the cell confluency reached 70%, the medium was discarded, washed twice with PBS, then digested with 400 μL trypsin for 2 min, and 3 mL of complete culture medium was used to terminate the digestion. After fu...

Embodiment 2

[0035] Pig placenta was nourished with DMEM:F12 medium containing 100nM and 10mM N-acetylcysteine ​​(98mL of DMEM:F12 in 100mL culture solution, 1mL of ITS and 1mL of penicillin / streptomycin double antibody solution) Layer cells, porcine large follicle granulosa cells, and porcine large follicle theca cells (wherein, porcine large follicle granulosa cells and theca cells were isolated from sow ovaries collected from slaughterhouses), cultured for 24 hours, and these cells were treated with N-acetyl-free DMEM with cysteine: F12 medium was used as blank control. After the culture was terminated, the cells were collected for cell counting. Three different replicates were designed for this experiment, each with 4 wells.

[0036] Carry out calculation by the cell counting method among the embodiment 1, the result obtained is: for the culture medium containing 100nM N-acetylcysteine, pig placental trophoblast cell proliferation reaches 1.6 times compared with blank control (P figur...

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Abstract

The present invention provides a new application of N-acetylcysteine, specifically, the application of N-acetylcysteine ​​in the preparation of pig placental trophoblast cell in vitro culture agent can be used to promote pig placental nourishment trophoblast cells in vitro, or inhibit the proliferation of porcine placental trophoblast cells in vitro.

Description

technical field [0001] The invention relates to the technical field of livestock breeding, in particular to the application of N-acetylcysteine ​​in in vitro culture of pig placenta trophoblast cells. Background technique [0002] The placental barrier is composed of trophoblast cells, capillary endothelial cells and basement membrane. It is a place for the exchange of nutrients and metabolites between the mother and the fetus, and it is also a natural barrier to prevent various pathogens from entering the fetus from the mother. Placental development is the result of proliferation and differentiation of trophoblast cells, and placental trophoblast cells are the key to the formation and development of placenta. [0003] N-acetylcysteine ​​(N-acetylcysteine, NAC) is a thiol compound, a precursor of the natural amino acid L-cysteine ​​and glutathione, and its molecular formula is C 5 h 9 NO 3 S, with a relative molecular weight of 163.2, is a compound containing active sulfh...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N5/073
CPCC12N5/0605C12N2500/32
Inventor 冯涛刘彦白佳桦许晓玲宋玉清肖霖力肖银霞
Owner BEIJING ACADEMY OF AGRICULTURE & FORESTRY SCIENCES