Cell cryopreservation liquid, application thereof and cryopreservation method of megakaryocyte progenitor cells

Abstract

The invention relates to the field of cell culture, in particular to cell cryopreservation liquid, application thereof and a cryopreservation method of megakaryocyte progenitor cells. The cell cryopreservation liquid comprises DMSO, fetal calf serum, dextran, trehalose and albumin. By means of the cell cryopreservation liquid, the cytoactive of the megakaryocyte progenitor cells can be well maintained during the cryopreservation period, and damage to cells in the cryopreservation and resuscitation processes is lowered. According to the cell cryopreservation liquid, application thereof and the cryopreservation method of the megakaryocyte progenitor cells, the cryopreservation and temperature reduction processes are softer, the cells are stored in liquid nitrogen after the temperature reduction process is conducted, the influence on the cytoactive is small, and damage to the cells in the cryopreservation process is lowered. It is indicated by experiments that the vigour of the resuscitated cells can reach 94% one month after the megakaryocyte progenitor cells are cryopreservated in the cryopreservation liquid, and multiplication activity is good, and the cell cryopreservation liquid is significantly superior to the prior art.

Description

technical field

[0001] The invention relates to the field of cell culture, in particular to a cell cryopreservation solution and its application and a method for cryopreservation of megakaryotic progenitor cells. Background technique

[0002] Megakaryocyte progenitor cell (megakaryocyte progenitor cell) is a kind of cell that makes people produce megakaryocyte, which is derived from bone marrow progenitor (CFU-GEMM), referred to as megakaryocyte progenitor cell. Megakaryotic progenitor cells exist in the IL-6R- cell population, while CD34+ cells can be divided into two cell subsets, expressing IL-6R and not expressing IL-6R, IL-6R+ cells can be stimulated to form granule-macrophages, Lymphocytes and granulocytes, while IL-6R- cells can form erythroid and megakaryocytes when given IL-6 / sIL-6R. Hematopoietic stem cells first differentiate into megakaryotic progenitor cells, also known as megakaryotic colony forming units (colonyforming unit-megakaryocyte, CFU-Meg). After the...

Examples

Embodiment 1

[0058] Prepare mother liquor:

[0059] Low-molecular-weight dextran mother liquor: purchased from Shanghai Changzheng Fumin Jinshan Pharmaceutical Co., Ltd., with a mass fraction of 10%.

[0060] Trehalose mother solution: fully dissolve 10mg trehalose powder in 0.5ml water

[0061] Human albumin mother solution: fully dissolve 120mg albumin in 3ml water

[0062] Add 0.5ml low-molecular dextran mother solution, 0.5ml trehalose mother solution, and 3ml human serum albumin mother solution to 5.5ml FBS in sequence, and finally add 0.5ml DMSO slowly to FBS, blow and mix gently, and store in a 4°C refrigerator .

Embodiment 2

[0064] Prepare mother liquor:

[0065] Low-molecular-weight dextran mother liquor: purchased from Shanghai Changzheng Fumin Jinshan Pharmaceutical Co., Ltd., with a mass fraction of 10%.

[0066] Trehalose mother solution: fully dissolve 10mg trehalose powder in 0.5ml water

[0067] Human albumin mother solution: fully dissolve 120mg albumin in 3ml water

[0068] Add 1ml low-molecular dextran mother solution, 0.5ml trehalose mother solution, and 3ml human serum albumin mother solution to 4.5ml FBS in sequence, and finally add 1ml DMSO slowly to FBS, mix gently by pipetting, and store in a 4°C refrigerator.

Embodiment 3

[0070] Prepare mother liquor:

[0071] Low-molecular-weight dextran mother liquor: purchased from Shanghai Changzheng Fumin Jinshan Pharmaceutical Co., Ltd., with a mass fraction of 10%.

[0072] Trehalose mother solution: fully dissolve 20mg trehalose powder in 1ml water

[0073] Human albumin mother solution: fully dissolve 80mg albumin in 2ml water

[0074] Add 1ml low-molecular dextran mother solution, 1ml trehalose mother solution, and 2ml human serum albumin mother solution to 5ml FBS in sequence, and finally add 1ml DMSO slowly to FBS, mix gently by pipetting, and store in a 4°C refrigerator.

technical field

[0001] The invention relates to the field of cell culture, in particular to a cell cryopreservation solution and its application and a method for cryopreservation of megakaryotic progenitor cells. Background technique

[0002] Megakaryocyte progenitor cell (megakaryocyte progenitor cell) is a kind of cell that makes people produce megakaryocyte, which is derived from bone marrow progenitor (CFU-GEMM), referred to as megakaryocyte progenitor cell. Megakaryotic progenitor cells exist in the IL-6R- cell population, while CD34+ cells can be divided into two cell subsets, expressing IL-6R and not expressing IL-6R, IL-6R+ cells can be stimulated to form granule-macrophages, Lymphocytes and granulocytes, while IL-6R- cells can form erythroid and megakaryocytes when given IL-6 / sIL-6R. Hematopoietic stem cells first differentiate into megakaryotic progenitor cells, also known as megakaryotic colony forming units (colonyforming unit-megakaryocyte, CFU-Meg). After the...

Examples

Embodiment 1

[0058] Prepare mother liquor:

[0059] Low-molecular-weight dextran mother liquor: purchased from Shanghai Changzheng Fumin Jinshan Pharmaceutical Co., Ltd., with a mass fraction of 10%.

[0060] Trehalose mother solution: fully dissolve 10mg trehalose powder in 0.5ml water

[0061] Human albumin mother solution: fully dissolve 120mg albumin in 3ml water

[0062] Add 0.5ml low-molecular dextran mother solution, 0.5ml trehalose mother solution, and 3ml human serum albumin mother solution to 5.5ml FBS in sequence, and finally add 0.5ml DMSO slowly to FBS, blow and mix gently, and store in a 4°C refrigerator .

Embodiment 2

[0064] Prepare mother liquor:

[0065] Low-molecular-weight dextran mother liquor: purchased from Shanghai Changzheng Fumin Jinshan Pharmaceutical Co., Ltd., with a mass fraction of 10%.

[0066] Trehalose mother solution: fully dissolve 10mg trehalose powder in 0.5ml water

[0067] Human albumin mother solution: fully dissolve 120mg albumin in 3ml water

[0068] Add 1ml low-molecular dextran mother solution, 0.5ml trehalose mother solution, and 3ml human serum albumin mother solution to 4.5ml FBS in sequence, and finally add 1ml DMSO slowly to FBS, mix gently by pipetting, and store in a 4°C refrigerator.

Embodiment 3

[0070] Prepare mother liquor:

[0071] Low-molecular-weight dextran mother liquor: purchased from Shanghai Changzheng Fumin Jinshan Pharmaceutical Co., Ltd., with a mass fraction of 10%.

[0072] Trehalose mother solution: fully dissolve 20mg trehalose powder in 1ml water

[0073] Human albumin mother solution: fully dissolve 80mg albumin in 2ml water

[0074] Add 1ml low-molecular dextran mother solution, 1ml trehalose mother solution, and 2ml human serum albumin mother solution to 5ml FBS in sequence, and finally add 1ml DMSO slowly to FBS, mix gently by pipetting, and store in a 4°C refrigerator.

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