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Cryopreservatio solution and cryopreservation method for human peripheral blood mononuclear cells

A human peripheral blood, nuclear cell technology, applied in the field of cell biology, can solve the problems of apoptosis, decreased cell activity, easy to change cytochemical and physical environment, etc., to achieve high cell activity, high amplification multiple, increase cell activity and recovery rate

Inactive Publication Date: 2020-03-31
FUJIAN YINFENG STEM CELL ENG +1
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  • Claims
  • Application Information

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Problems solved by technology

However, this cryopreservation method easily changes the chemical and physical environment of the cells during the cryopreservation process, and can easily cause biological damage to the cells.
Ice crystals are easy to form during cryopreservation, resulting in reduced cell activity after recovery, a large number of dead cells, and poor cell expansion, cell senescence and apoptosis

Method used

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  • Cryopreservatio solution and cryopreservation method for human peripheral blood mononuclear cells
  • Cryopreservatio solution and cryopreservation method for human peripheral blood mononuclear cells
  • Cryopreservatio solution and cryopreservation method for human peripheral blood mononuclear cells

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Embodiment 1

[0019] Reagents: lymphocyte separation medium (GE, 1714403); serum-free medium (Corning, KBM581); NK amplification reagent (Hangzhou Zhongying Biomedical Technology Co., Ltd.); interleukin 2 (Shandong Quangang Pharmaceutical Co., Ltd., 20170917) .

[0020] 1. Acquisition of human peripheral blood mononuclear cells:

[0021] (1) Collect 140ml of peripheral blood from a healthy donor, wipe the outer surface of the blood collection tube with a 75% alcohol-free dust-free cloth, and then separate the peripheral blood mononuclear cells.

[0022] (2) Centrifuge the peripheral blood at 2000rmp for 8min; divide 15ml / tube of lymphocyte separation solution into 50ml centrifuge tubes.

[0023] (3) After the peripheral blood is centrifuged, draw the upper plasma into a 50ml centrifuge tube, inactivate the plasma at 56°C for 30 minutes, centrifuge, and take the supernatant, which is autologous plasma. Dilute the blood cells in the lower layer with normal saline, the volume ratio of blood ...

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Abstract

The invention discloses a cryopreservation solution and a cryopreservation method for human peripheral blood mononuclear cells, and belongs to the technical field of cell biology. The cryopreservationsolution comprises the following components: 8% of dimethyl sulfoxide, 70% of autologous plasma, 10% of hydroxyethyl starch and 12% of human serum albumin. The specific cryopreservation procedure comprises the following steps: a, waiting at 4 DEG C until the product is put in; b, cooling to 0 DEG C at a speed of 1 DEG C / min, and keeping for 5 minutes; c, cooling to -10 DEG C at a speed of 1 DEG C / min, and keeping for 5 minutes; d, cooling to -45 DEG C at a speed of 1 DEG C / min, and keeping for 20 minutes; e, cooling to -90 DEG C at a speed of 5 DEG C / min, and keeping for 5 minutes; and f, ending. According to the cryopreservation solution and the cryopreservation method, the cell activity and the recovery rate after cryopreservation recovery can be remarkably increased, high-quality NK cells with high purity, high cell activity and high amplification factor are induced and cultured, and a favorable guarantee is provided for clinical treatment of the NK cells.

Description

technical field [0001] The invention belongs to the technical field of cell biology, and in particular relates to a cryopreservation solution and a cryopreservation method for human peripheral blood mononuclear cells. Background technique [0002] Peripheral blood cells include red blood cells, white blood cells, and platelets, and peripheral blood mononuclear cells (PBMC) are cells with a single nucleus in peripheral blood, which are a type of white blood cells, including lymphocytes and monocytes. PBMC is a blood cell with a round nucleus. In both myeloid and lymphoid lineages, there are related cells that make up PBMC. It is composed of different proportions of lymphocytes (T cells, B cells and NK cells), dendritic A cell population consisting of cells and monocytes. PBMC is a key component of the immune system, so it is widely used in many fields such as immunology, infectious diseases and transplantation therapy. [0003] Natural killer cells (NK) are a type of lympho...

Claims

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Application Information

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IPC IPC(8): A01N1/02
CPCA01N1/0221A01N1/0284
Inventor 肖桂清唐燕程郑梦盈苏爱盟谢燕萍
Owner FUJIAN YINFENG STEM CELL ENG
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