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Monocyte culture solution

A culture medium and nuclear cell technology, applied in the biological field, can solve the problems of high price, unclear composition of Sigma culture medium, unfavorable promotion, etc., achieve the effect of less usage, avoid potential risks, and maintain group characteristics

Active Publication Date: 2012-10-24
SHENZHEN BEIKE BIOTECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0002] Mononuclear cells are a mixed population of various cells. The Sigma medium prepared with the combination of Sigma basal medium and growth factors can better expand MNC cells, but the composition of the Sigma medium is not clear and the price is expensive, which is not conducive to wide application. to promote

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment

[0018] Embodiment 1 The preparation of mononuclear cell culture medium.

[0019] The mononuclear cell culture fluid provided by the present invention includes basal culture fluid, cell growth auxiliary components and growth factors, specifically: IMDM culture fluid + 10 μg / mL recombinant human insulin + 20 μg / mL transferrin + 100 μmol / L β-mercaptoethanol + 5~10ng / mL recombinant human thrombopoietin+5~10ng / mL recombinant human stem cell growth factor+5~10ng / mL rhFlt-3 ligand+5~10ng / mL interleukin 6+5~10ng / mL interleukin 3+5~10ng / mL recombinant human granulocyte-macrophage colony-stimulating factor.

[0020] In order to facilitate the comparison of cell culture effects, a variety of different culture media were prepared as follows:

[0021] Auto6-10: IMDM medium + 10μg / mL recombinant human insulin + 20μg / mL transferrin + 100μmol / L β-mercaptoethanol + 10ng / mL recombinant human thrombopoietin + 10ng / mL recombinant human stem cell growth factor + 10ng / mL rhFlt-3 ligand+10ng / mL in...

Embodiment 3

[0049] Example 3 Detection of colony-forming ability of mononuclear cells.

[0050] Mononuclear cells are a mixed population of cells that contain hematopoietic stem and progenitor cells. Under appropriate culture conditions, hematopoietic stem cells and progenitor cells will differentiate into erythroid and myeloid cell colonies, producing erythroid colonies (CFU-E), burst erythroid colonies (BFU-E), granulocyte colonies (CFU-G), Colonies of macrophages (CFU-M), colonies of granulosa-macrophages (CFU-GM) and large mixed colonies (CFU-GEMM).

[0051] The present invention uses the cell colony forming ability detection kit of R&D Systems, Inc., the article number is HSC005. Operate according to the product manual, divide into 5 time periods of 0, 3, 6, 9 and 12 days, take 4×10 respectively 4 Resuspend cells in 300 μL of cell suspension, add to 3 mL of methylcellulose enhanced medium pre-thawed at 4 degrees or room temperature, mix well, and divide 1.1 mL / 35 mm plate into two ...

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Abstract

The invention provides monocyte culture solution and belongs to the field of biotechnology. The monocyte culture solution comprises Iscove's Modified Dulbecco's Medium (IMDM) basic culture solution, recombinant human insulin, transferrin, beta-mercaptoethanol, recombinant human thrombopoietin, recombinant human stem cell growth factor, rhFlt-3ligand, interleukin6, interleukin3 and recombinant human granulocyte-macrophage colony stimulating factor. The monocyte culture solution provided by the invention is clear in components, and can realize the quick amplification of monocytes and better keep the group property of the monocytes; meanwhile, material and coat are saved. The solution is mainly used for culturing monocytes.

Description

technical field [0001] The invention belongs to the field of biotechnology, in particular to a mononuclear cell culture solution. Background technique [0002] Mononuclear cells are a mixed population of various cells. The Sigma medium prepared with the combination of Sigma basal medium and growth factors can better expand MNC cells, but the composition of the Sigma medium is unclear and expensive, which is not conducive to wide application. promote. Contents of the invention [0003] In order to solve the problems existing in the prior art (the composition of the culture medium is unclear and the cost is high), the present invention provides a mononuclear cell culture medium with clear composition, which can be used for the rapid expansion of mononuclear cells and saves the production cost. [0004] A mononuclear cell culture medium, including IMDM culture medium, recombinant human insulin, transferrin, β-mercaptoethanol, recombinant human thrombopoietin, recombinant hum...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N5/078
Inventor 黄昕华汪照静朱姣莲李陶姜舒胡祥
Owner SHENZHEN BEIKE BIOTECH
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