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A culture medium for human peripheral blood lymphocytes

A technology of lymphocytes and human peripheral blood, applied in the field of cell culture medium, which can solve the problems of complex components and difficult control, and achieve good culture effect and good culture effect

Active Publication Date: 2022-03-29
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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  • A culture medium for human peripheral blood lymphocytes
  • A culture medium for human peripheral blood lymphocytes
  • A culture medium for human peripheral blood lymphocytes

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0026] Embodiment 1 A kind of 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, lectin 54mg / L.

[0028] The preparation method is as follows:

[0029] 1) Weigh 10.4g of RPMI 1640 medium powder and dissolve it in 1L of ultrapure water, add 2.0g of sodium bicarbonate and 2.38g of 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 formulation requirements: gentamicin sulfate, heparin sodium, mercaptoethanol, glutamine, calcium ions, iron ions, titanium ions, and mix well.

[0032] 4) After mixing the above solution, filter it with a 0.22 μm filter membrane, then add the phytohemagglu...

Embodiment 2

[0033] Embodiment 2 A kind of 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, lectin 72mg / L.

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

Embodiment 3

[0036] Embodiment 3 A kind of 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, lectin 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 medium and discloses a human peripheral blood lymphocyte culture medium. The medium of the present invention mainly comprises the following components: RPMI 1640 medium powder, sodium bicarbonate, HEPES free acid, gentamicin sulfate, heparin sodium, mercaptoethanol, glutamine, calcium ion, iron ion, titanium ion, plant blood Lectin. The culture medium of the present invention may contain serum or protein components, or may be free of serum and protein components, and it mainly utilizes certain specific metal ions to promote cell proliferation. The culture medium of the invention has clear components, stable performance and low cost, can effectively promote the proliferation and division of lymphoblasts in human whole blood, and has a relatively high lymphocyte transformation rate 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 formula of human peripheral blood lymphocyte culture medium and a preparation method thereof. Background technique [0002] Lymphocytes in human peripheral blood are almost in the GI phase (G0 phase), and generally do not continue to divide. If conA, lipopolysaccharide (LPS), and interleukin When IL-2, phytohemagglutinin (PHA) and other mitogens and antigenic substances are released, small lymphocytes will be stimulated by the inducer to transform into lymphoblasts, and then enter mitosis. After short-term cell culture, colchicine solution treatment, hypotonicity and fixation, some metaphase cells of mitosis can be obtained. This method has been widely used in clinical medicine, virology, pharmacology, genetic toxicity and so on. [0003] Causes known to affect the growth of human peripheral blood lymphocytes are: [0004] 1) Virus or microbial infect...

Claims

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Application Information

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Patent Type & Authority Patents(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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