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Aminotic cell culture medium, aminotic cell culture method, and application of culture medium

An amniotic fluid cell and culture method technology, which is applied to cell culture active agents, embryonic cells, animal cells, etc., can solve the problems that the product is not widely used in clinical practice and the technology is not mature enough, and achieves the effect of speeding up the culture and improving the success rate.

Inactive Publication Date: 2018-05-22
SHENZHEN LONGGANG DISTRICT MATERUITY & CHILD HEALTHCARE HOSPITAL
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

At present, many domestic prenatal diagnosis institutions use imported German AMNIOPAN amniotic fluid medium and American Gibico amniotic fluid medium. Domestic Hunan Xiangya Medical Gene Technology Co., Ltd., Chongqing and Guangzhou also have several companies conducting research on this technology, but the technology Not yet mature enough, the product has not been widely used clinically

Method used

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  • Aminotic cell culture medium, aminotic cell culture method, and application of culture medium
  • Aminotic cell culture medium, aminotic cell culture method, and application of culture medium
  • Aminotic cell culture medium, aminotic cell culture method, and application of culture medium

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0027] Please refer to Figure 1-9 , Embodiment 1 of the present invention is:

[0028] 1. Preparation of medium components

[0029] The basic components of the amniotic fluid cell medium are: basic culture medium, bovine serum, cell growth factors, double antibodies, etc., but the amount of cells in the amniotic fluid itself is small, and most of them are aging cells. Therefore, compared with peripheral blood, amniotic fluid cell culture is difficult. It is much larger, and the preparation of the medium is the key to the success or failure of amniotic fluid cell culture. The preparation process must ensure the quality of the raw materials used, and the specific formula of each component must also be verified through experiments, and the activity of each biological component must be ensured during the preparation and use process, and the most important point is to ensure that the preparation and use process is sterile , once contaminated, the entire experimental process will...

Embodiment 2

[0040] The amniocyte culture medium of the present embodiment is different from the first embodiment only in "the volume fraction of fetal bovine serum is 4-15%", and the others are the same as the first embodiment;

[0041] The culture conditions of the amniocytes of this embodiment are as follows: take 1 mL of primary amniocyte samples at the age of 22 weeks, inoculate them in 5 mL of optimized different culture media for comparative culture, culture them in culture media with different fetal bovine serum ratios, and replace them after 7 days. After 11 days, observe the growth of amniotic fluid cells and harvest the cells. Colchicine was added 1.5 hours before terminating the culture to arrest as many amniocytes as possible in metaphase.

Embodiment 3

[0043] The amniocyte culture medium of the present embodiment is different from the first embodiment only in "the volume fraction of fetal bovine serum is 4-15%", and the others are the same as the first embodiment;

[0044]The culture conditions of the amniocytes in this example are as follows: take 0.7 mL of a 20-week-old primary amniocyte sample, inoculate it in 5 mL of optimized different culture media for comparative culture, and culture it in media with different fetal bovine serum ratios for 6.5 days After changing the medium, observe the growth of amniotic fluid cells after 10.5 days and harvest the cells. Colchicine was added 1.5 hours before terminating the culture to arrest as many amniocytes as possible in metaphase.

[0045] test test

[0046] The amniocyte culture medium prepared in Examples 1 to 3 is used as a test example, wherein the data in Example 1 is used as a representative example, and Example 1 is Test Example 1; meanwhile, the U.S. Gibico amniocyte cu...

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Abstract

The invention relates to an aminotic cell culture medium, which is prepared from a basic culture medium, and further is prepared from the following component: transferrin, sodium selenite, insulin, triiodothyronine, glucagon, progesterone, hydrocortisone, testosterone, estradiol, a basic fibroblast growth factor, vitamin E, vitamin C, antibiotic and fetal calf serum. The invention also relates toapplication of the aminotic cell culture medium in antenatal diagnosis of aminotic cell growth and an aminotic cell culture method. The aminotic cell culture medium provided by the invention has theadvantages of capability of promoting aminotic cell proliferation and excellent quality of the aminotic cell culture medium.

Description

technical field [0001] The invention relates to cell culture technology, in particular to an amniotic fluid cell culture medium, a method for culturing amniotic fluid cells and the application of the culture medium. Background technique [0002] China is a country with a large population in the world, and it is also a country with a high incidence of birth defects and disabilities. Birth defects are not only a serious public health problem, but also a social problem that affects economic development and people's normal life. Birth defects and disabilities have not only increasingly become important issues that affect the quality of the population, but also impose a heavy economic burden on families and society. Prenatal / prenatal diagnosis Karyotype analysis is the best way to effectively reduce birth defects. At present, major hospitals in China have carried out the karyotype analysis technology of amniotic fluid and villi. The quality of the amniotic fluid medium is the k...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N5/073C12Q1/02
CPCC12N5/0605C12N2500/12C12N2500/24C12N2500/30C12N2500/32C12N2500/35C12N2500/38C12N2500/84C12N2501/115C12N2501/33C12N2501/335G01N33/5044
Inventor 魏凤香万新红米占英胡亮罗小金温丽娟王艳春
Owner SHENZHEN LONGGANG DISTRICT MATERUITY & CHILD HEALTHCARE HOSPITAL
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