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In vitro culture medium and in vitro culture system for culturing human embryos

An in vitro culture and culture medium technology, applied in the direction of embryonic cells, culture process, tissue culture, etc., can solve the problems of high price, medium change in the middle, unsupported embryo growth and development, etc., to reduce the impact, improve the culture effect, reduce the The effect of the action steps

Active Publication Date: 2018-07-06
SHENZHEN VITAVITRO BIOTECH CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

At present, the domestic culture system generally has the following defects: (1) Sequential culture medium is mostly used, and the medium needs to be changed midway during the culture process; (2) Most of the domestic purchases of imported reagents are expensive and the procurement cycle is long; (3) The whole process of embryo culture The bottom of the dish is placed flat, and there is no microenvironment to support the growth and development of embryos; for this reason, a new culture system is necessary to solve the above problems

Method used

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  • In vitro culture medium and in vitro culture system for culturing human embryos
  • In vitro culture medium and in vitro culture system for culturing human embryos
  • In vitro culture medium and in vitro culture system for culturing human embryos

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0019] [Example 1] Embryo culture fluid formula and preparation method:

[0020] A. Culture medium formula:

[0021]

[0022]

[0023] B. Preparation method:

[0024] a. Sterilize the equipment used in the preparation at high temperature and dry it for later use. The whole preparation process is carried out in a clean workshop that meets the requirements;

[0025] b. Add the weighed ultrapure water, and then add sodium chloride, potassium chloride, potassium dihydrogen phosphate, calcium chloride dihydrate, magnesium sulfate dihydrate, Disodium EDTA, essential amino acids and non-essential amino acids, glucose, inositol, sodium citrate, taurine, sodium pyruvate, sodium lactate, start the agitator until the solid matter is completely dissolved, and make base liquid A;

[0026] c. Add the accurately weighed gentamicin sulfate and phenol red of each formula in the formula table to the above base liquid A, and finally add sodium bicarbonate, start the agitator until the so...

Embodiment 2

[0029] [Example 2] Detection of Embryo Culture Solution

[0030] pH detection

[0031] Take an appropriate amount of the sample to be tested and measure it three times with a pH meter, and take the average value of the three data obtained as the result, and it is considered qualified if it is 7.3±0.1.

[0032] Osmotic pressure test

[0033] After the osmometer is calibrated, take 50μl of the sample to be tested and put it into the test tube to start the test. When the value is stable, read the data, measure 3 data according to the above method, take the average as the result, and it is considered qualified if it is 257-273mOsm / kg .

[0034] Bacterial endotoxin detection

[0035] According to the requirements of the 2015 edition of the Pharmacopoeia of the People's Republic of China, the test is carried out by the limulus reagent gel method, and the result is ≤0.25EU / ml, which is considered qualified.

[0036] Cytotoxicity

[0037] Tested according to the provisions of GB / ...

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Abstract

The invention provides an in vitro culture medium for culturing human embryos, and a preparation method of the in vitro culture medium, belonging to the field of human assisted reproduction. The in vitro culture medium is mainly prepared from the following components: 0.01-0.05g / L of gentamicin sulfate, 0.005-0.01g / L of phenol red, 0-0.01mM / L of disodium ethylenediaminetetraacetate, 90-116mM / L ofsodium chloride, 2.5-10.0 mM / L of potassium chloride, 0-1.8 mM / L of calcium chloride dihydrate, 0-7.16 mM / L of potassium dihydrogen phosphate, 0.2-1.51M / L of magnesium sulfate dihydrate, 20-25 mM / L of sodium bicarbonate, 0.1-7.27 mM / L of sodium pyruvate, 0-20 mM / L of sodium lactate, 0-3.25 mM / L of glucose, 0-1.0 mM / L of sodium citrate, 0-1.0mM / L of L-alanyl-L-glutamine, 0-2.7mM / L of inositol, 0.05-5mM / L of taurine, 5g / L of human serum protein, essential amino acids and non-essential amino acids.

Description

technical field [0001] The invention belongs to the field of human assisted reproduction, and specifically relates to an in vitro culture technology of embryo cells, including a culture medium capable of supporting the development of early human embryos to blastocysts and a culture system of a culture device. Background technique [0002] The first human in vitro fertilization (IVF) baby was born in 1978. Since then, assisted reproduction has flourished, and egg single sperm injection (ICSI), preimplantation genetic diagnosis (PGD), egg or embryo cryopreservation and other technologies have been born one after another. The earth has promoted the development of the assisted reproductive industry. [0003] Embryo culture fluid provides pre-implantation embryos with the nutrients needed for development, and provides an in vitro environment for clinically required specific period embryo transfer (ET) or pre-implantation genetic diagnosis (PGD). Indispensable reagents in experim...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N5/073C12M1/00C12M1/22
CPCC12M23/10C12M23/34C12N5/0603C12N2500/05C12N2500/32C12N2500/33C12N2500/34C12N2500/35C12N2501/998
Inventor 林小贞
Owner SHENZHEN VITAVITRO BIOTECH CO LTD
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