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Granular cell removing solution and preparation method thereof

A technology for granulating cells and removing liquid, which is applied in cell dissociation methods, biochemical equipment and methods, biological preparations for removing bad cells, etc., can solve the problems of lack of antioxidants and metal ion chelating agents, etc. damage reduction effect

Active Publication Date: 2020-10-16
SHENZHEN VITAVITRO BIOTECH CO LTD
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  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The base liquid of the existing granule cell removal liquid products is a simple salt solution, which lacks antioxidants and metal ion chelating agents

Method used

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  • Granular cell removing solution and preparation method thereof
  • Granular cell removing solution and preparation method thereof
  • Granular cell removing solution and preparation method thereof

Examples

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Embodiment

[0025] 1. Formula and preparation method of egg washing solution:

[0026] A. Granule cell removal liquid formula:

[0027]

[0028]

[0029] B. Preparation method:

[0030] a. Weigh all the solid components according to the formula of the above granular cell removal solution and place them separately;

[0031] b. Add and dissolve all components except gentamycin sulfate, hyaluronidase and human serum albumin in ultrapure water according to solid first and then liquid;

[0032] c, adding gentamicin sulfate to the solution obtained in step b, and finally adding hyaluronidase and dissolving to obtain a basic buffer;

[0033] e. Add human serum albumin in proportion to the above basic buffer and mix well;

[0034] f. Measure and adjust the pH value of the above-mentioned base solution at 7.3±0.1, and the osmotic pressure at 260-290mOsm / kg;

[0035] g. Filter the solution through a 0.22 μm membrane filter in a 100-grade isolator, divide it into packages, seal it, and sto...

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Abstract

The invention belongs to the technical field of human assisted reproduction, and relates to a granular cell removing solution of human oocyte. The solution is prepared by dissolving a proper amount ofEDTA and taurine in a basic salt solution containing a granular cell removing enzyme. The content of the EDTA is 0.005 to 0.015 mM / L. The content of the taurine is 0.05 to 0.15 mM / L. The granular cell removing enzyme is hyaluronidase, and the content of the granular cell removing enzyme is 50-90 IU / mL. The basic salt solution comprises the following components: 90.08 to 101.5 mM / L of sodium chloride, 4.41 to 5.50 mM / L of potassium chloride, 0.50 to 2.20 mM / L of calcium chloride dihydrate, 0.30 to 0.44 mM / L of potassium dihydrogen phosphate, 0.15 to 2.00 mM / L of magnesium sulfate heptahydrate,3.50 to 14.50 mM / L of sodium bicarbonate, 10.5 to 21.5 mM / L of 4-hydroxyethylpiperazine ethane sulfonic acid (HEPES), 0.01 to 0.05 g / L of gentamicin sulfate, 0.50 to 2.88 mM / L of glucose, 0.31 to 0.35 mM / L of sodium pyruvate, 10.5 to 21.50 mM / L of L-sodium lactate and 5 to 10 g / L of human serum albumin.

Description

technical field [0001] The invention belongs to the technical field of human assisted reproduction, and relates to a liquid for removing granulosa cells of human oocytes and a preparation method thereof. [0002] technical background [0003] Before intracytoplasmic sperm injection or egg freezing, oocytes need to be prepared, mainly by using special granulosa cell digestion enzymes to remove granulosa cells around the oocytes. The granulosa cell removal solution is made by adding hyaluronidase to the buffered saline solution, and it is used for assisted reproduction for the digestion of granulosa cells around the egg. The base liquid of the existing granule cell removal liquid product is a simple salt solution, which lacks anti-oxidation and metal ion chelating agents. Contents of the invention [0004] The object of the present invention is to provide a granulosa cell removal liquid containing antioxidant and chelating agent, which can effectively digest and loosen ovum ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N5/00C12N5/075
CPCC12N5/0081C12N5/0609C12N2509/00
Inventor 林小贞
Owner SHENZHEN VITAVITRO BIOTECH CO LTD
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