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Bone marrow stem cell preservation and transportation method

A technology of bone marrow stem cells and transportation method, applied in the biological field, can solve the problems of cytotoxicity, unfavorable clinical application, and the effect of stem cells is not very good, and achieve the effect of high safety and low uncertainty of raw materials

Inactive Publication Date: 2016-05-04
北京弘润天源基因生物技术有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

In the prior art, the effect of using lymphocyte separation medium to separate stem cells is not very good, and the conventional Ficoll400 separation medium has certain toxicity to cells, which is not conducive to clinical application

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0025] A method for storing and transporting bone marrow stem cells, comprising the following steps:

[0026] 1) Obtain red bone marrow according to clinical routine bone marrow aspiration;

[0027] 2) Add 5 times the volume of PBS to the bone marrow and gently mix and dilute, centrifuge at 400g for 5 minutes, discard the supernatant and fat layer, and obtain bone marrow cells; add an equal volume of PBS to the bone marrow cells and mix gently to obtain bone marrow cell suspension liquid;

[0028] 3) Add the separation solution containing trehalose and ultra-low viscosity sodium alginate into the centrifuge tube, spread the bone marrow cell suspension on the separation solution, and centrifuge at 8-25°C, 200g-800g, 10-30min, Absorb buffy coat cells to obtain bone marrow stem cells;

[0029] 4) The bone marrow stem cells were mixed in equal proportions with the cryopreservation solution containing 2% trehalose + 8% glycerol + 10% DMSO + 18% autologous plasma + 62% normal sali...

Embodiment 2

[0034] A method for storing and transporting bone marrow stem cells, comprising the following steps:

[0035] 1) Obtain red bone marrow according to clinical routine bone marrow aspiration;

[0036]2) Add 5 times the volume of PBS to the bone marrow and gently mix and dilute, centrifuge at 400g for 5 minutes, discard the supernatant and fat layer, and obtain bone marrow cells; add an equal volume of PBS to the bone marrow cells and mix gently to obtain bone marrow cell suspension liquid;

[0037] 3) Add the separation solution containing trehalose and ultra-low viscosity sodium alginate into the centrifuge tube, spread the bone marrow cell suspension on the separation solution, and centrifuge at 8-25°C, 200g-800g, 10-30min, Absorb buffy coat cells to obtain bone marrow stem cells;

[0038] 4) The bone marrow stem cells were mixed in equal proportions with the cryopreservation solution containing 2% trehalose + 8% glycerol + 10% DMSO + 18% autologous plasma + 62% normal salin...

Embodiment 3

[0043] A method for storing and transporting bone marrow stem cells, comprising the following steps:

[0044] 1) Obtain red bone marrow according to clinical routine bone marrow aspiration;

[0045] 2) Add 5 times the volume of PBS to the bone marrow and gently mix and dilute, centrifuge at 400g for 5 minutes, discard the supernatant and fat layer, and obtain bone marrow cells; add an equal volume of PBS to the bone marrow cells and mix gently to obtain bone marrow cell suspension liquid;

[0046] 3) Add the separation solution containing trehalose and ultra-low viscosity sodium alginate into the centrifuge tube, spread the bone marrow cell suspension on the separation solution, and centrifuge at 8-25°C, 200g-800g, 10-30min, Absorb buffy coat cells to obtain bone marrow stem cells;

[0047] 4) The bone marrow stem cells were mixed in equal proportions with the cryopreservation solution containing 2% trehalose + 8% glycerol + 10% DMSO + 18% autologous plasma + 62% normal sali...

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Abstract

The invention discloses a bone marrow stem cell preservation and transportation method and belongs to the technical field of biology. The method comprises the following steps that red marrow is obtained according to the clinical routine; the marrow is added into 5-fold PBS, centrifugation is carried out, and marrow cells are obtained; PBS is added into the marrow cells, and the mixture is lightly stirred to form suspension; separation liquid containing trehalose is added into a centrifugal pipe, the suspension is flatly laid on the separation liquid, centrifugation is carried out, and albuginea layer cells are absorbed; the cells and a freezing medium are mixed in equal proportion to be subpackaged in a freezing pipe; the mixture is frozen at -80 DEG C, and then transferred into liquid nitrogen to be frozen; the freezing pipe is taken out of the liquid nitrogen and immediately placed into a constant-temperature water bath tank to be lightly waggled to be rethawed; the rethawed cell suspension cell is transferred to the centrifugal pipe containing normal saline at 3-7 DEG C, mixed to be uniform and centrifuged, and bone marrow stem cells are collected; the bone marrow stem cells are added into a preservation solution containing 0.5% trehalose, and transported to a using unit through a heat insulation box with the constant temperature of 3-7 DEG C within 24 h. By means of the method, the activity of the cells is kept in the transportation process.

Description

technical field [0001] The invention relates to the field of biotechnology, in particular to a method for preserving and transporting bone marrow stem cells. Background technique [0002] Bone marrow is an important immune organ of the human body, containing a large number of immune cells, including stem cells, lymphocytes, etc. During clinical application, a common problem is that it is difficult to ensure a sufficient number of stem cells. At present, stem cells are mainly preserved by cryopreservation technology, revived and induced to cultivate various immune cells when needed. However, in the process of separation and cryopreservation, stem cells will be damaged or even toxic, especially in the process of cryopreservation, which will significantly change the thermodynamic, chemical and physical environment of the cells, accompanied by the risk of biological damage. [0003] On the other hand, bone marrow contains a lot of fat, which affects the separation effect and ca...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N5/0775C12N5/0789A01N1/02
CPCA01N1/0221A01N1/0236C12N5/0647C12N5/0663
Inventor 宫喜魁李若鲲马艳马艳玲郝丽敏
Owner 北京弘润天源基因生物技术有限公司
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