Human peripheral blood lymphocyte medium and preparation method thereof

A lymphocyte and culture medium technology, applied in the field of human peripheral blood lymphocyte culture medium and its preparation, can solve the problems of lack of nutrients, easy to be infected by viruses or microorganisms, unstable pH, etc., and achieve good reagent stability, inhibition Microorganisms and viruses, the effect of high mitotic index of lymphocytes

Inactive Publication Date: 2017-03-22
叶宗耀
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0012] Aiming at the problems that the existing cell culture medium is easy to be infected by viruses or microorganisms, lack of nutr...

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0027] Embodiment 1 A kind of human peripheral blood lymphocyte culture medium

[0028] A culture medium for human peripheral blood lymphocytes, consisting of the following components and their concentrations: RPMI1640 medium 55g / L, calf serum 1.5g / L, collagen 4.76g / L, sodium hyaluronate 3.75g / L, pancreatic Glucagon 1.43mg / L, thyrotropin releasing hormone 6.8μg / L, total glycosides of paeony 22.56mg / L, linoleic acid 2.75g / L, butylenediamine 0.32g / L, hydrated ferric gluconate 3.5mg / L, glutathione 4.25μg / L, polylysine 3.15mg / L.

[0029] The preparation method is divided into the following steps:

[0030] (1) Accurately weigh the dry powder of RPMI1640 medium and place it in a sterilized container, add ultrapure water to dilute, then stir and mix to obtain solution I;

[0031] (2) adopting the pasteurization method to dissolve the bovine serum and add it to the solution I after the low-temperature treatment to obtain the solution II;

[0032] (3) Collagen, sodium hyaluronate, ...

Embodiment 2

[0033] Embodiment 2 A kind of human peripheral blood lymphocyte culture medium

[0034] A culture medium for human peripheral blood lymphocytes, consisting of the following components and their concentrations: RPMI1640 medium 56g / L, calf serum 2g / L, collagen 5g / L, sodium hyaluronate 4.5g / L, glucagon Thyrotropin 1.5mg / L, thyrotropin 7μg / L, total glycosides of paeony 30mg / L, linoleic acid 3g / L, butanediamine 0.5g / L, hydrated ferric gluconate 5mg / L, glutathione Peptide 5μg / L, polylysine 3.5mg / L.

[0035] The preparation method of the above medium is similar to that of Example 1.

Embodiment 3

[0036] Embodiment 3 A kind of human peripheral blood lymphocyte culture medium

[0037] A culture medium for human peripheral blood lymphocytes, consisting of the following components and their concentrations: RPMI1640 medium 50g / L, calf serum 1g / L, collagen 4g / L, sodium hyaluronate 3g / L, glucagon 0.1mg / L, thyrotropin 5μg / L, total glucosides of paeony 15mg / L, linoleic acid 1g / L, butanediamine 0.1g / L, hydrated ferric gluconate 1mg / L, glutathione 0.1μg / L, polylysine 0.5mg / L.

[0038] The preparation method of the above medium is similar to that of Example 1.

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PUM

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Abstract

The invention belongs to the technical field of cell mediums, and in particular relates to a human peripheral blood lymphocyte medium and a preparation method thereof. The medium provided by the invention mainly consists of the following components: an RPMI1640 medium, fetal bovine serum, collagen, sodium hyaluronate, glucagon, thyrotropin releasing hormone, total glucosides of paeony, linoleic acid, butanediamine, gluconic acid iron salt hydrate, glutathione and polylysine. The medium provided by the invention can be used for effectively inhibiting microorganisms and viruses, and the medium is good in reagent stability and high in transformation rate of cultured lymphocytes and lymphocyte division index.

Description

technical field [0001] The invention belongs to the technical field of cell culture medium, and in particular relates to a human peripheral blood lymphocyte culture medium and a preparation method thereof. Background technique [0002] The small lymphocytes in human peripheral blood are almost in the GI phase (GO phase), and generally do not divide anymore. When phytohemagglutinin PHA is added to the culture medium, these small lymphocytes are stimulated to transform into lymphocytes. Mother cells then enter mitosis, so after short-term culture, colchicine treatment, hypotonicity and fixation, some mitotic cells can be obtained. This method has been widely used in clinical medicine, virology, pharmacology, genetic toxicity and so on. [0003] At present, the reasons that affect the growth of human peripheral blood lymphocytes are: [0004] (1) Virus or microbial infection: When cells are cultured in vitro, compared with in vivo cells lose their defense against microorganis...

Claims

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Application Information

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IPC IPC(8): C12N5/0781C12N5/0783
CPCC12N5/0635C12N5/0636C12N2500/24C12N2500/30C12N2500/36C12N2500/46C12N2500/50C12N2500/76C12N2501/335C12N2501/375C12N2501/905
Inventor 叶宗耀
Owner 叶宗耀
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