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Human oocyte in-vitro mature culture solution and preparing method and culture method thereof

A technology for in vitro maturation and cultivation of human oocytes, applied in the medical field, can solve the problems of complicated medium preparation methods and cultivation methods, high cost, low in vitro maturation rate of human oocytes, etc. Productivity and productivity, effects on increased cleavage and blastocyst formation rates

Active Publication Date: 2019-04-16
周桦 +2
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0004] The object of the present invention is to make up for the deficiencies in the prior art, providing a human oocyte in vitro maturation culture medium and its preparation method and culture method, which has Improve the in vitro maturation rate of human oocytes, simplify the medium preparation method and culture method, reduce the cost of purchasing commercial in vitro maturation medium, and solve the low in vitro maturation rate of human oocytes, complicated medium preparation methods and culture methods , the problem of high cost of purchasing commercial in vitro maturation medium

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0023] A human oocyte maturation medium in vitro, characterized in that: 70 parts of 75IU / L rFSH, 60 parts of 150IU / L rHCG, 50 parts of 20% SPS, 30 parts of 2mg / L EGF, 20 parts of 25mmol / L sodium pyruvate And basic cell culture medium Qiunn's1029 150 parts.

[0024] Its preparation method is as follows:

[0025] S1, first put 150 parts of the basic cell culture solution Qiunn's 1029 into the reactor, and add 70 parts of 75IU / L rFSH and 60 parts of 150IU / L rHCG, then start the reaction kettle, make it fully mixed, and then leave it for 30 minutes to let it Fully integrated;

[0026] S2, sequentially add 50 parts of 20% SPS, 30 parts of 2mg / L EGF and 20 parts of 25mmol / L sodium pyruvate to the inside of the reactor, and start the reactor at the same time, keep the temperature at 5°C, mix for 30 minutes to make it fully mixed uniform;

[0027] S3, put the semi-finished product obtained in step S2 into a medical refrigerator, keep the temperature inside the refrigerator at 2-8°...

Embodiment 2

[0034] A human oocyte maturation medium in vitro, characterized in that: 80 parts of 75IU / L rFSH, 65 parts of 150IU / L rHCG, 53 parts of 20% SPS, 30 parts of 2mg / L EGF, 23 parts of 25mmol / L sodium pyruvate And basic cell culture medium Qiunn's1029 170 parts.

[0035] Its preparation method is as follows:

[0036] S1, first put 170 parts of the basic cell culture solution Qiun's 1029 into the reactor, and add 80 parts of 75IU / L rFSH and 65 parts of 150IU / L rHCG, then start the reactor, make it fully mixed, and then leave it for 30 minutes to let it Fully integrated;

[0037] S2, sequentially add 53 parts of 20% SPS, 30 parts of 2mg / L EGF and 23 parts of 25mmol / L sodium pyruvate into the reactor, and start the reactor at the same time, keep the temperature at 5°C, mix for 30 minutes to make it fully mixed uniform;

[0038] S3, put the semi-finished product obtained in step S2 into a medical refrigerator, keep the temperature inside the refrigerator at 2-8°C, and the preparatio...

Embodiment 3

[0045] A human oocyte maturation medium in vitro, characterized in that: 95 parts of 75IU / L rFSH, 70 parts of 150IU / L rHCG, 55 parts of 20% SPS, 40 parts of 2mg / L EGF, 25 parts of 25mmol / L sodium pyruvate And basic cell culture medium Qiunn's1029 200 parts.

[0046] Its preparation method is as follows:

[0047] S1, first put 200 parts of the basic cell culture solution Qiunn's 1029 into the reactor, and add 95 parts of 75IU / L rFSH and 70 parts of 150IU / L rHCG, then start the reaction kettle, make it fully mixed, and then leave it for 30 minutes to make it Fully integrated;

[0048] S2, sequentially add 55 parts of 20% SPS, 40 parts of 2mg / L EGF and 25 parts of 25mmol / L sodium pyruvate to the interior of the reactor, and start the reactor at the same time, keep the temperature at 5°C, and mix for 30 minutes to make it fully mixed. uniform;

[0049] S3, put the semi-finished product obtained in step S2 into a medical refrigerator, keep the temperature inside the refrigerator...

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Abstract

The invention discloses a human oocyte in-vitro mature culture solution and a preparing method and culture method thereof, and relates to the technical field of medicines. The formula of the culture solution includes, by mass, 70-95 parts of 75IU / L rFSH, 60-70 parts of 150IU / L rHCG, 50-55 parts of 20% SPS, 30-40 parts of EGF with the concentration of 2 mg / L, 20-25 parts of sodium pyruvate with theconcentration of 25 mmol / L and 150-200 parts of a basic cell culture solution Qiunn's 1029. According to the human oocyte in-vitro mature culture solution and the preparing method and culture methodthereof, the cost for purchasing a commercial in-vitro mature culture medium can be effectively reduced; in the culture medium, the human blastocyst culture solution Qiunn's 1029 serves as the basic culture medium, the culture medium is safe and free of toxic and side effects, the meiosis of immature oocytes, especially oocytes in a third-stage cumulus oophorus and oocyte complex, can be promoted,the in-vitro development ability of this type of cells is improved, the maturity of cytoplasm is promoted, the after-cell-maturity cleavage rate and blastocyst development rate are increased, and theembryo quality is further improved.

Description

technical field [0001] The invention relates to the field of medical technology, in particular to a human oocyte maturation culture solution in vitro and a preparation method and a culture method thereof. Background technique [0002] Oocytes refer to the oogonia that undergo meiosis during oogenesis, and are divided into primary oocytes, secondary oocytes, and mature oocytes. Production, the first meiosis and the product of the second meiosis. [0003] As an important part of assisted reproductive technology, extracellular maturation of oocytes can not only improve the survival efficiency of oocytes, reduce the occurrence of ovarian hyperstimulation syndrome, but also provide more options for fertility preservation of egg-donating women. The existing culture medium preparation method is expensive to purchase commercial in vitro maturation medium, the in vitro maturation rate of human oocytes is low, and the medium preparation and culture methods are complicated. For this r...

Claims

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Application Information

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IPC IPC(8): C12N5/075
CPCC12N5/0609C12N2501/31C12N2500/30C12N2501/11
Inventor 周桦
Owner 周桦
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