A serum-free human peripheral blood lymphocyte culture medium
A lymphocyte and culture medium technology, applied in the field of biological preparations, can solve problems such as complex serum components, increased risk of spreading diseases, fluctuations in serum quality, etc., achieve high lymphocyte conversion rate and lymphocyte division index, and reduce transmission of diseases Risk, effect of reducing culture medium cost
Active Publication Date: 2018-02-27
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
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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 preparation, in particular to a lymphocyte culture medium. It includes lectin, basal medium and serum substitute, the lectin is L-type phytohemagglutinin, the basal medium is RPMI1640 basal medium, and the serum substitute includes bovine serum albumin, recombinant human insulin , ferric citrate and amino acid mother liquor. The serum-free human peripheral blood lymphocyte culture medium of the present invention does not use animal or human serum, which can not only reduce the cost of the culture medium, but also reduce the risk of spreading diseases; the serum-free human peripheral blood lymphocyte culture medium of the present invention is used The components are determined, which avoids the problem of complex components of traditional serum and quality differences between different batches, which lead to fluctuations in the culture effect; the cell culture medium provided by the invention has a high lymphocyte conversion rate and lymphocyte division index, low reagent cost and high quality. Stable, cultured cells can be used for clinical diagnosis of 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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IPC IPC(8): C12N5/0781C12N5/0783
Inventor 周海军吴家波
Owner 广州和能生物科技有限公司