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Oocyte activating solution and application thereof

A technology of oocytes and solutions, applied in the field of assisted reproduction, can solve the problems of low activation success rate of chemical activators, achieve enhanced embryo development potential, improve oocyte fertilization rate and blastocyst formation rate, and improve embryo development quality Effect

Active Publication Date: 2020-11-17
成都艾伟孚生物科技有限公司
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  • Abstract
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AI Technical Summary

Problems solved by technology

[0006] However, the activation success rate of existing chemical activators is low. Tian Jianhui et al. found through research that porcine oocytes were activated by ionomycin and then treated with 6-DMP for 5.5 hours. The activation effect was better, but the blastocyst rate was about 25.4% (Tian Jianhui, CAI Yuan, LIU Guoshi, et al.Effects of Chemical Activation on Parthenogenetic Development of Porcine Oocytes in Vitro[J].Chinese Agricultural Sciences,2005,038(005):1029-1033.)

Method used

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  • Oocyte activating solution and application thereof
  • Oocyte activating solution and application thereof

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Embodiment 5

[0035] The preparation method of embodiment 5 oocyte activation liquid

[0036] The preparation method of described oocyte activation liquid, comprises the following steps:

[0037] S1. Under the condition of grade 100, weigh the reduced glutathione, heparin, ionomycin, A23187, puromycin, 6-DMAP, SrCl according to the ratio 2 , phosphatidylserine, sodium chloride, potassium chloride, calcium chloride, magnesium sulfate, potassium dihydrogen phosphate, sodium bicarbonate, glucose, amino acids or their salts, alanyl glutamine, sodium lactate, sodium pyruvate, bovine Sulfonic acid, HEPES, MOPS, human serum albumin, gentamycin sulfate are added to water and mixed uniformly to obtain solution I;

[0038] S2. Weigh the cycloheximide CHX, phorbol ester PMA, IP3, and paclitaxel according to the proportion, and add them to ethanol and mix evenly. The concentration of each component in absolute ethanol is 1000 times of the final working concentration according to the proportion. , to ...

Embodiment 6

[0043] Example 6 A method for improving the fertilization rate of mammalian oocytoplasmic sperm microinjection

[0044] The methods include:

[0045] After intracytoplasmic sperm microinjection for 10 min, the oocytes were placed in the balanced oocyte activation solution at a temperature of 37°C and O 2 The concentration is 5%, CO 2 Concentration of 6% of the environment under the conditions of treatment for 20min, with the equilibrium step culture medium Wash 5 times, then move directly into one-step medium In, at a temperature of 37°C, O 2 The concentration is 5%, CO 2 The micro-drop culture was carried out for 6 hours under the environmental condition of 6% concentration.

[0046] The oocyte activation solution consists of the following components and their contents:

[0047] Ionomycin 1.5mg / L, calcium ionophore A23187 8mg / L, puromycin 80μg / L, 6-DMAP 30mg / L, cycloheximide CHX 5mg / L, phorbol ester PMA 40μg / L, muscle triphosphate Alcohol 10μg / L, SrCl 2 1.8g / L, pa...

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Abstract

The invention belongs to the technical field of assisted reproduction, and relates to an oocyte activating solution and application thereof. The oocyte activating solution comprises 1.1 to 1.8 mg / L ofionomycin, 6 to 12 mg / L of calcium ion carrier A23187, 63 to 94 mug / L of puromycin, 19 to 43 mg / L of 6DMAP, 3.4 to 6.6 mg / L of actinone CHX, 28 to 53 mug / L of phorbol PMA, 7.8 to 12.7 mug / L of inositol triphosphate, 1.5 to 2.4 g / L of SrCl2, 16 to 25 mg / L of heparin, and basic culture medium. The oocyte activating solution can effectively activate oocytes, and is suitable for activating the oocytes after ICSI in an in-vitro fertilization assisted reproduction technology.

Description

technical field [0001] The invention belongs to the technical field of assisted reproduction, and relates to an oocyte activation solution and an application thereof. Background technique [0002] Before fertilization, mammalian mature oocyte development stays in the meiosis MII stage, and only when fertilization or fertilization-like signal stimulation occurs, the oocyte restarts the activation process, leaves this stage, and enters the next stage, The second meiosis is resumed, two pronuclei are formed, the second polar body is discharged, and the embryo continues to develop. This stage is called oocyte activation. [0003] The activation of oocytes is a complex signal transduction process induced by the endoplasmic reticulum and inositol triphosphate IP3 signaling pathways that lead to an increase in the concentration of calcium ions in the cytoplasm. During conventional in vitro fertilization (IVF), intracytoplasmic Ca 2+ The increase in concentration occurs within a f...

Claims

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Application Information

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IPC IPC(8): C12N5/075C12N5/073
CPCC12N5/0609C12N5/0604C12N2501/91C12N2501/999C12N2500/12C12N2500/14C12N2500/16C12N2500/34C12N2500/32C12N2500/30C12N2500/35C12N2500/33C12N2501/998Y02A40/81
Inventor 余裕炉叶林刘洪君严飞戴甄
Owner 成都艾伟孚生物科技有限公司
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