Oocyte activating physical method and fertilization potential evaluation index thereof

An oocyte and evaluation index technology, applied in the field of embryo engineering, can solve the problems of cell damage, excessive energy, poor repeatability, etc.

Active Publication Date: 2017-11-03
PEOPLES HOSPITAL PEKING UNIV +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0006] These methods can improve the activation efficiency of oocytes to a certain extent, but the operation of chemical and mechanical methods is complicated, the repeatability is poor, and the activation efficiency and embryonic development potential need to be improved
However, the traditional electrical activation method has too much energy, which is easy to damage the cells.

Method used

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  • Oocyte activating physical method and fertilization potential evaluation index thereof
  • Oocyte activating physical method and fertilization potential evaluation index thereof
  • Oocyte activating physical method and fertilization potential evaluation index thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0025] Example 1: Activation of rabbit oocytes

[0026] 1. Experimental animals

[0027] Sexually mature female Japanese big-eared rabbits aged 6-15 months and weighing 2.5-3.5 kg were purchased from Beijing Weitong Lihua Laboratory Animal Technology Co., Ltd.

[0028] 2. Experimental reagents

[0029] Pregnant Horse Serum (PMSG Sigma)

[0030] Human chorionic gonadotropin (hCG Sigma)

[0031] Plasma Substitute (SSS Irvine Scientific)

[0032] Hyaluronidase (Sigma)

[0033] Modified Human Fallopian Tube Fluid (mHTF Irvine Scientific)

[0034] Human tubal fluid (HTF Irvine Scientific)

[0035] 3. Oocyte Preparation and Culture

[0036] First, the neck skin of female rabbits was disinfected with iodine cotton balls, and PMSG (150IU / rabbit) was subcutaneously injected into the neck. After 48 hours, the ear edge was sterilized with alcohol cotton balls, and hCG (75IU / a) was injected into the ear vein for superovulation. 24h after the injection of hCG, the female rabbits w...

Embodiment 2

[0044] Example 2: Induction of calcium ion oscillations in mouse oocytes

[0045] 1. Experimental animals

[0046] Sexually mature female Kunming mice aged 6-8 weeks were purchased from Beijing Weitong Lihua Laboratory Animal Technology Co., Ltd.

[0047] 2. Experimental reagents

[0048] Pregnant Horse Serum (PMSG Sigma)

[0049]Human chorionic gonadotropin (hCG Sigma)

[0050] Plasma Substitute (SSS Irvine Scientific)

[0051] Hyaluronidase (Sigma)

[0052] Modified Human Fallopian Tube Fluid (mHTF Irvine Scientific)

[0053] Human tubal fluid (HTF Irvine Scientific)

[0054] Fluo-4AM / Calcium Fluorescent Probe (Dojindo Laboratories)

[0055] 3. Oocyte Preparation and Culture

[0056] PMSG (10IU / mice) was injected intraperitoneally to female mice, and hCG (10IU / mice) was injected intraperitoneally 48 hours later for superovulation. After 15-19 hours of hCG injection, the mice were sacrificed by cervical dislocation, and the fallopian tubes were exposed and removed un...

Embodiment 3

[0061] Example 3: Evaluation of the fertilization potential of A23187-activated mouse oocytes

[0062] 1. Experimental animals

[0063] Sexually mature female Kunming mice aged 6-8 weeks were purchased from Beijing Weitong Lihua Laboratory Animal Technology Co., Ltd.

[0064] 2. Experimental reagents

[0065] Pregnant Horse Serum (PMSG Sigma)

[0066] Human chorionic gonadotropin (hCG Sigma)

[0067] Plasma Substitute (SSS Irvine Scientific)

[0068] Hyaluronidase (Sigma)

[0069] Modified Human Fallopian Tube Fluid (mHTF Irvine Scientific)

[0070] Human tubal fluid (HTF Irvine Scientific)

[0071] Fluo-4AM / Calcium Fluorescent Probe (Dojindo Laboratories)

[0072] Calcium ionophore A23187 (A23187Sigma)

[0073] 3. Oocyte Preparation

[0074] PMSG (10IU / mice) was injected intraperitoneally to female mice, and hCG (10IU / mice) was injected intraperitoneally 48 hours later for superovulation. After 15-19 hours of hCG injection, the mice were sacrificed by cervical dislo...

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PUM

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Abstract

The invention relates to an oocyte activating physical method and a fertilization potential evaluation index thereof. An electronic pulse sequence is used for activating the oocyte; the calcium oscillation feature formed by calcium ion concentration change in the endochylema detected in a confocal microscopy is used as an oocyte fertilization potential evaluation index.

Description

Technical field: [0001] The invention relates to the field of embryo engineering, in particular to a physical method for activating oocytes in the field of assisted reproduction, and an evaluation index for evaluating the fertilization potential of oocytes activated by sperm or artificial assistance methods. technical background [0002] Today, about 10%-16% of couples worldwide face infertility problems. Although the birth of assisted reproductive technology has helped many infertile patients, the phenomenon of fertilization failure is still common. In the field of assisted reproduction, intracytoplasmic sperm injection (ICSI) is the most widely used technique. It uses a microscope operating system to manually inject a single sperm into the oocyte to fertilize it. The fertilization rate was significantly increased to 70%. However, there are still a considerable number of patients who are unable to fertilize after ICSI. Studies have found that insufficient oocyte activatio...

Claims

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Application Information

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IPC IPC(8): C12N5/075C12N13/00G01N21/64G01N21/63
CPCC12N5/0609C12N13/00G01N21/63G01N21/6486
Inventor 鹿群刘佳卉梁蓉国晋菘王建六王凯乐董飞宏张珏方竞
Owner PEOPLES HOSPITAL PEKING UNIV
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