Application of NAD signal channel agonist in in-vitro embryo culture

A signaling pathway and in vitro culture technology, applied in the field of biomedicine, can solve the problems of early embryonic developmental block and low efficiency of "test-tube baby" treatment

Pending Publication Date: 2022-05-31
童国庆
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The prominent problem caused is: 40%-60% of human embryos have early developmental arrest, and most embryos stop developing in the early stages of development, and then die, resulting in low efficiency of "test-tube baby" treatment, especially for women older than 35 years old and young women (less than 35 years old) with premature decline in ovarian function

Method used

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  • Application of NAD signal channel agonist in in-vitro embryo culture
  • Application of NAD signal channel agonist in in-vitro embryo culture
  • Application of NAD signal channel agonist in in-vitro embryo culture

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0049] Example 1 Ovum maturation culture

[0050] The female subjects were subjected to ovulation induction, and the size of the follicles was monitored. When the diameter of the follicles reached 6-12 mm, the immature eggs were obtained by puncturing by B-ultrasound.

[0051] On the day of egg retrieval, prepare the 3037 insemination dish according to the "IVF-ET laboratory procedures and laboratory preparation" routine, and mark the patient's name and medical record number on the lid and bottom of the dish, and put it into a three-gas incubator for balance. At the beginning of egg picking, the laboratory egg picking operator and the nurse jointly check the three certificates of the patient. During egg retrieval, the man enters the sperm collection room to collect semen. Before egg retrieval, a certain amount of round-bottomed Falcon 2001 test tubes were placed in a 37°C thermostatic test tube rack to preheat. Prepare 5-10 egg-picking dishes (Falcon 3003) and place them in ...

Embodiment 2

[0052] Example 2 Washing Human Sperm

[0053] After traditional testicular sperm acquisition, it was placed in Modified-HTF (Irvine Scientific company) containing 1mM NR, and then incubated in a 5% CO2 box for 10 to 24 hours. Sperm motility was significantly improved.

Embodiment 3

[0054] Example 3 Sperm freezing and thawing

[0055] Each 100mL human semen cryoprotectant contains the following substances:

[0056] Glucose 1.5g, trisodium citrate dihydrate 1.3g, glycerol 20ml, NR 1mM, egg yolk 20ml, dimethyl sulfoxide 0.1ml, and the balance is double distilled water.

[0057] After mixing human semen cryoprotectant and semen according to a volume ratio of 1:3, add a test tube and use a program-controlled cooling device to freeze. The freezing procedure is set as follows: (1) Equilibrate at 20°C for 5 minutes; 2°C, freezing rate -1°C / min; (3) from -2°C to -30°C, freezing rate -7.2°C / min; (4) from -30°C to -90°C, freezing rate -30°C / min.

[0058] After freezing, the frozen human sperm was thawed, and the motility of the frozen-thawed sperm and the recovery rate of sperm after freezing and thawing were evaluated, using the sperm percentage as an indicator of forward motility. The inventors found that adding NR to the freezing and thawing recovery solutio...

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Abstract

The invention provides a method for culturing embryonic cells in vitro, and according to the embodiment of the invention, the method comprises the following steps: (i) in vitro culturing fertilized egg cells or derivatives thereof in the presence of an NAD (Nicotinamide Adenine Dinucleotide) signaling pathway agonist. In the culture process of fertilized egg cells or derivatives thereof, the NAD signal channel agonist is added, so that the quality, such as development potential, of embryos cultured in vitro can be effectively improved.

Description

technical field [0001] The invention relates to the field of biomedicine, and in particular, the invention relates to the application of an NAD signaling pathway agonist in in vitro embryo culture. Background technique [0002] In human in vitro fertilization and embryo transfer treatment (commonly known as "test tube baby"), early immature eggs, mature eggs, sperm, fertilized eggs, and early embryos need to be incubated in a specific culture system. The current physical environment such as culture temperature, humidity, and oxygen content is basically perfect and constant. However, although egg in vitro maturation medium, fertilized egg culture medium and early embryo culture medium are commercially produced and applied, they are far from perfect and mature. meet medical expectations. The prominent problems caused are: 40%-60% of human embryos have early developmental block, most embryos stop developing in the early stage of development, and then die, resulting in low effi...

Claims

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Application Information

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IPC IPC(8): C12N5/073C12N5/076C12N5/075
CPCC12N5/0604C12N2506/04C12N2501/999C12N2500/38C12N2500/40C12N2501/40
Inventor 童国庆张翔
Owner 童国庆
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