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A low serum amniotic fluid culture medium

A technology of amniotic fluid cells and medium, applied in the field of cell biology, can solve the problems of not being able to generate enough mitotic cells, and achieve the effects of large-scale application, increase of clone formation rate and clone growth speed, and cost reduction

Active Publication Date: 2017-07-21
广州和能生物科技有限公司
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Problems solved by technology

When the planted amniotic fluid sample is in poor condition, it is impossible to generate enough mitotic cells in a short period of time, which still has a certain gap with the needs of clinical testing units

Method used

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  • A low serum amniotic fluid culture medium
  • A low serum amniotic fluid culture medium
  • A low serum amniotic fluid culture medium

Examples

Experimental program
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Effect test

Embodiment

[0019] Embodiment: preparation low serum amniotic fluid cell culture medium of the present invention (code name HN-1):

[0020] Take 5.0g of dry powder of RPMI1640 medium and 5.1g of dry powder of αMEM medium, dissolve them in 950ml of ultrapure water, and add NaHCO 3 2.0g, ferric citrate 5mg, recombinant human insulin (Insulin) 10mg, recombinant human basic fibroblast growth factor (bFGF) 30ug, recombinant human epidermal growth factor (EGF) 10ug, bovine serum albumin (BSA) 2.0g , Hydrocortisone (hydrocortisone) 2nMol, α-tocopherol (Alpha Tocopherol) 1mg, ethanolamine 1mg, oleic acid, linoleic acid 1mg, then add Tween 80 (Tween 80) 0.5ml, fetal bovine serum (FBS) 10ml, after mixing, add ultrapure water to make up to 1000ml, which is the complete medium of HN-1 amniocytes. Aseptically filter and aliquot, store at -20°C for later use.

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Abstract

The invention discloses a low serum amniotic fluid cell culture medium. On the basis of mixing a basal culture medium RPMI 1640 and a basal culture medium alpha MEN according to the proportion of one to one, the components of insulin, a basic fibroblast growth factor (bFGF), an epidermal growth factor (EGF), bovine serum albumin (BSA), hydrocortisone, alpha tocopherol, NaHCO3, ferric citrate, tween 80, ethanol amine, oleic acid, linoleic acid and fetal calf serum are added, wherein addition of fetal calf serum content accounts for not more than 1% of the total volume of the low serum amniotic fluid cell culture medium. According to the low serum amniotic fluid cell culture medium, compared with an existing public technology, the serum content and the cost are reduced, the effect is significantly improved, colony forming efficiency and colony growth speed are increased obviously, in a short time, a great number of aminotic cells in a division phase can be obtained through culture, and the requirements of clinical diagnoses and scientific research can be met.

Description

technical field [0001] The invention belongs to the field of cell biology, in particular to a low-serum amniotic fluid cell culture medium. Background technique [0002] Prenatal diagnosis is an important part of eugenics. It is based on genetic counseling. Through prenatal testing of fetal health, early measures can be taken for children with serious genetic diseases and congenital deformities to reduce the accumulation of harmful genes in the population. , in order to reduce the birth of children and improve the quality of the population, so it is of great significance. Prenatal diagnosis currently mainly includes villous tissue sampling in the first trimester, amniotic fluid and umbilical cord blood in the second trimester, separation of fetal cells from peripheral blood of pregnant women, and preimplantation diagnosis, but amniotic fluid cell examination in the second trimester is still the most important diagnostic method. [0003] Suitable objects for amniocentesis ex...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N5/073
Inventor 周海军吴家波
Owner 广州和能生物科技有限公司
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