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Serum-free human peripheral blood lymphocyte culture medium

A lymphocyte and peripheral blood technology, applied in the field of lymphocyte culture medium, can solve the problems of complex serum composition, increase the risk of spreading diseases, and fluctuate serum quality, achieve high lymphocyte conversion rate and lymphocyte division index, and reduce transmission. Risk of disease, effect of reducing medium cost

Active Publication Date: 2015-08-12
广州和能生物科技有限公司
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Problems solved by technology

However, the disadvantages of using animal serum are also obvious: due to the complex composition of serum, there are not only growth-promoting substances, but also potential growth-inhibiting substances; and the quality fluctuations of different batches of serum lead to batch-to-batch differences in the effect of culture medium cells
[0003] The technical problem to be solved by the present invention is that most of the existing peripheral blood lymphocyte culture medium uses animal or human serum as an additive, which not only costs a lot, but also increases the risk of spreading diseases, and at the same time, there are problems of quality differences between different batches

Method used

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  • Serum-free human peripheral blood lymphocyte culture medium
  • Serum-free human peripheral blood lymphocyte culture medium
  • Serum-free human peripheral blood lymphocyte culture medium

Examples

Experimental program
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Effect test

Embodiment 1

[0022] (1) Preparation of 50X amino acid mother liquor

[0023] The L-type amino acids used are all from Sinopharm Group (BR grade), and the specific preparation of 1L amino acid mother liquor is as follows:

[0024] Weigh 300mg of L-glutamic acid, 100mg of L-glycine, and 400mg of L-aspartic acid respectively, add them into 500mL of ultrapure water, stir well, add ultrapure water to make up to 1000mL, mix well, and pack in 4 Store at ℃ for later use.

[0025] (2) The preparation method of PHA-L

[0026] (1) PHA crude extract extraction:

[0027] 1. Material: red kidney beans (fresh, no mold, bright color).

[0028] 2. Crush the qualified red kidney beans and pass through a 100-mesh sieve to make red kidney beans powder. Take a certain amount of red kidney bean powder, soak it with twice the amount of normal saline, and soak it in the refrigerator for 12 hours. The samples were centrifuged (20° C., 10000 rpm, 15 min), and the supernatant was collected. Add ethanol to the ...

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Abstract

The invention relates to a biological agent and particularly relates to a lymphocyte culture medium. The lymphocyte culture medium comprises lectin, a basal culture medium and a serum substitution, wherein the lectin is L-type phytohemagglutinin; the basal culture medium is an RPMI1640 basal culture medium; and the serum substitution comprises bovine serum albumin, recombinant human insulin, ferric citrate and an amino acid mother solution. By adopting the serum-free human peripheral blood lymphocyte culture medium, the animal or human serum is not used so that the cost of the culture medium is reduced and the disease dissemination risk is also reduced. The components of the serum-free human peripheral blood lymphocyte culture medium are determined so that the problems of complex traditional serum components, quality differences in different batches and fluctuated culture effect can be avoided. The culture medium provided by the invention is high in lymphocyte conversion rate and lymphocyte metaphase index, low in reagent cost and stable in quality, and the cultured cell can be used for clinical diagnosis of the karyotype analysis.

Description

technical field [0001] The invention relates to a biological preparation, in particular to a lymphocyte culture medium. Background technique [0002] With the development of cytogenetics, karyotype analysis of peripheral blood lymphocytes has become a routine detection item in prenatal diagnosis. Lymphocyte culture medium is the main material of this test project, and its product quality directly affects the success rate and accuracy of the test. Conventional peripheral blood lymphocyte culture medium is to add a certain amount of animal serum (such as bovine serum) on the basis of basal medium (such as RPMI1640) to provide the necessary substances for cell growth and proliferation. However, the disadvantages of using animal serum are also obvious: due to the complex composition of serum, there are not only growth-promoting substances, but also potential growth-inhibiting substances; and the quality fluctuations of different batches of serum lead to batch-to-batch differenc...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N5/0781C12N5/0783
Inventor 周海军吴家波张邦骤
Owner 广州和能生物科技有限公司
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