Human peripheral blood lymphocyte culture medium

A lymphocyte and human peripheral blood technology, applied in the field of cell culture medium, can solve the problems of complex components and difficult control, and achieve the effects of high transformation rate and division index, good stability of reagents, and precise control of the amount of addition

Active Publication Date: 2020-01-10
HANGZHOU GENE META MEDICAL DEVICE CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0010] Aiming at the problem that the existing cell culture medium contains serum, protein and other biological extracts, resulting in complicated and difficult-to-control components, the present invention provides a culture medium for human peripheral blood lymphocytes and a preparation method thereof

Method used

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  • Human peripheral blood lymphocyte culture medium
  • Human peripheral blood lymphocyte culture medium
  • Human peripheral blood lymphocyte culture medium

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0026] Example 1 A human peripheral blood lymphocyte culture medium

[0027] RPMI 1640 medium 10.4g / L, sodium bicarbonate 2.0g / L, HEPES free acid 2.38g / L, bovine serum 5%, gentamicin sulfate 50μg / mL, heparin sodium 6.9U / mL, mercaptoethanol 7.15μmol / L, glutamine 0.25mmol / L, calcium ion 0.8mmol / L, iron ion 1.86μmol / L, titanium ion 15μmol / L, phytohemagglutinin 54mg / L.

[0028] The preparation method is as follows:

[0029] 1) Weigh 10.4g RPMI 1640 medium powder and dissolve it in 1L ultrapure water, add 2.0g sodium bicarbonate, 2.38g HEPES free acid, and mix well.

[0030] 2) Add 5% bovine serum of the total volume and mix well.

[0031] 3) Add the following substances to make the final concentration meet the formula requirements: gentamicin sulfate, heparin sodium, mercaptoethanol, glutamine, calcium ion, iron ion, titanium ion, and mix well.

[0032] 4) After mixing the above solution, filter it with a 0.22 μm filter membrane, then add the lectin required by the formula under aseptic co...

Embodiment 2

[0033] Example 2 A human peripheral blood lymphocyte culture medium

[0034] RPMI 1640 medium 10.4g / L, sodium bicarbonate 2.0g / L, HEPES free acid 2.38g / L, bovine serum albumin 0.5%, gentamicin sulfate 50μg / mL, heparin sodium 7.2U / mL, mercaptoethanol 7.5μmol / L, glutamine 0.2mmol / L, calcium ion 0.9mmol / L, iron ion 1.9μmol / L, titanium ion 15μmol / L, phytohemagglutinin 72mg / L.

[0035] The preparation method is similar to Example 1.

Embodiment 3

[0036] Example 3 A human peripheral blood lymphocyte culture medium

[0037] RPMI 1640 medium 10.4g / L, sodium bicarbonate 2.0g / L, HEPES free acid 2.38g / L, gentamicin sulfate 50μg / mL, heparin sodium 6.9U / mL, mercaptoethanol 7.15μmol / L, glutamine Amide 0.25mmol / L, calcium ion 0.8mmol / L, iron ion 1.86μmol / L, titanium ion 20.8μmol / L, phytohemagglutinin 54mg / L.

[0038] The preparation method is similar to Example 1.

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Abstract

The invention belongs to the technical field of cell culture mediums, and discloses a human peripheral blood lymphocyte culture medium. The human peripheral blood lymphocyte culture medium mainly comprises the following components: RPMI 1640 culture medium powder, sodium bicarbonate, HEPES free acid, gentamycin sulphate, heparin sodium, mercaptoethanol, glutamine, calcium ions, iron ions, titaniumions and phytohemagglutinin. The human peripheral blood lymphocyte culture medium can contain or be free of serum or protein components, and the culture medium mainly uses certain specific metal ionsto promote cell proliferation. According to the culture medium, the composition is clear, the performance is stable, the cost is low, proliferation and division of lymphoblasts in human whole blood can be effectively promoted, and the high lymphocyte transformation rate and a high lymphocyte division index are achieved.

Description

Technical field [0001] The invention belongs to the technical field of cell culture media, and specifically relates to a formula of a human peripheral blood lymphocyte culture medium and a preparation method thereof. Background technique [0002] Lymphocytes in human peripheral blood are almost all in the GI phase (G0 phase). Under normal circumstances, they will not continue to divide. If concanavalin (ConA), lipopolysaccharide (LPS), and leukocyte media are added to the medium When mitogens and antigenic substances such as IL-2 (IL-2) and phytohemagglutinin (PHA), small lymphocytes will be stimulated by inducers to transform into lymphoblasts, and then enter mitosis. After short-term cell culture, colchicine solution treatment, hypotonicity and fixation, some cells in metaphase can be obtained. This method has been widely used in clinical medicine, virology, pharmacology, and genetic toxicology. [0003] The known reasons that affect the growth of human peripheral blood lymphoc...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N5/078
CPCC12N5/0634C12N2500/12C12N2500/84C12N2500/44C12N2500/14C12N2500/24C12N2500/22C12N2501/998
Inventor 谭鸿浩王强
Owner HANGZHOU GENE META MEDICAL DEVICE CO LTD
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