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Cultivation method for activating human memory B cells into plasma cells in vitro

A technology for plasmablasts and culture methods, which is applied in the field of in vitro activation of human memory B cell plasmablasts, can solve the problems of low activation efficiency, low activation quality, and high cost, and achieve high activation efficiency and activation quality. The effect of small quantity and low cost

Inactive Publication Date: 2016-12-14
NANCHANG UNIV
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Problems solved by technology

[0004] Although the above combinations have successfully activated memory B cells, they more or less have the following disadvantages: the activation efficiency is not high, the activation quality is low, the operation is complicated, and the cost is expensive

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  • Cultivation method for activating human memory B cells into plasma cells in vitro

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Embodiment Construction

[0019] 1. Acquisition of memory B cells: collect whole blood from volunteers in a sterile environment, separate human peripheral blood mononuclear cells from fresh anticoagulated whole blood with lymphocyte separation medium, and then use human memory B cells magnetically The bead sorting kit separates memory B cells from peripheral blood mononuclear cells. Table 1 shows the number of human peripheral blood mononuclear cells; Table 2 shows the number of B lymphocytes; Table 3 shows the number of memory B lymphocytes.

[0020] 2. In vitro activation culture of memory B cells: the sorted memory B cells were inoculated into 96-well cell culture plates at the density of 0 cells / well, 5 cells / well, and 20 cells / well, and IMDM with 10% FBS was used as the basis Culture medium containing 30ng / mL of IL-2 and 60ng / mL of IL-21 and 5000cGy of 60 3T3-CD40L cells inactivated by Co irradiation were used as feeder cells at 10,000 / well at 37°C, 5% CO 2 Cultured under conditions for 13 days, ...

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Abstract

The invention discloses a cultivation method for activating human memory B cells into plasma cells in vitro. According to the method, irradiated 3T3-CD40L cells serve as feeder layer cells, IMDM of 10% FBS serves as basis culture solution, IL-2 and IL-21 serve as growth factors, and the memory B cells are induced in vitro to activate into the plasma cells and secrete antibodies. The cultivation method is simple to operate, low in cost, fewer in needed memory B cells and high in activation efficiency and activation quality.

Description

technical field [0001] The invention relates to a method for culturing plasmablasts of activated human memory B cells in vitro. Background technique [0002] Memory B cells are high-affinity cells retained by clonal elimination after the initial immune response. They are important immune cells in the body and play a very important role in humoral immunity. Protection plays a key role. The long-term immune memory mechanism of memory B cells plays an important guiding role in clinical treatment, vaccine development and fully human monoclonal antibody drug development. Due to the small number of plasma cells in human peripheral blood lymphocytes, the extraction is difficult. Even if a small amount can be extracted, the extraction method is cumbersome, high cost, and low acquisition rate, so it is not suitable for promotion. Therefore, the research on the activation of peripheral blood memory B cells in vitro has become a hotspot. [0003] In the 1980s, researchers used human...

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Application Information

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IPC IPC(8): C12N5/0781G01N33/53
CPCC12N5/0635C12N2501/2302C12N2501/2321C12N2502/13G01N33/53
Inventor 胡萍魏强辛洪波何晶晶
Owner NANCHANG UNIV
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