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Hematopoietic stem cell cryopreservation method

A technology of hematopoietic stem cells and cryopreservation method is applied in the field of cryopreservation of hematopoietic stem cells, which can solve the problems of differences in stem cell preservation effect and preservation time, complicated operations, and large differences in the use methods of protective agents, so as to improve the use effect of cryopreservation and reduce Degree of degradation or inactivation, effect of physical property stabilization

Pending Publication Date: 2021-10-29
合肥滴碧云生物科技有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

At present, the protective agents used for low-temperature preservation of stem cells at -80°C include dimethyl sulfoxide, CP-1, hydroxyethyl starch, RPMI1640 medium, human serum albumin, etc. The formulations and methods of use of different protective agents are quite different , There are also differences in the preservation effect and preservation time of stem cells. The survival rate of most preserved cells is about 80%. It is difficult to guarantee the safety of stem cells. Some cryopreservation methods also have the disadvantage of complicated operation.

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0059] The hematopoietic stem cell cryopreservation method of the present embodiment comprises the following steps:

[0060] (1) Pretreatment: Prepare hematopoietic stem cells and suspend them with normal saline;

[0061] (2) Enzymatic hydrolysis treatment: transfer the hematopoietic stem cells pretreated in step (1) to an enzymatic hydrolysis tank, perform enzymatic hydrolysis treatment, and then perform centrifugation treatment to obtain cell sediment;

[0062] (3) Suspension treatment: add the cell sediment after enzymatic hydrolysis treatment in step (2) to the suspension for treatment;

[0063] (4) Cryopreservation treatment: Suspend the hematopoietic stem cells treated in step (3), centrifuge and take out the cell pellet again, add stabilizing solution, and place in a -80°C refrigerator.

[0064] In the method for cryopreserving hematopoietic stem cells described above,

[0065] The hematopoietic stem cells described in step (1) were purchased from Beijing Zeping Techn...

Embodiment 2

[0093] The hematopoietic stem cell cryopreservation method of the present embodiment comprises the following steps:

[0094] (1) Pretreatment: Prepare hematopoietic stem cells and suspend them with normal saline;

[0095] (2) Enzymatic hydrolysis treatment: transfer the hematopoietic stem cells pretreated in step (1) to an enzymatic hydrolysis tank, perform enzymatic hydrolysis treatment, and then perform centrifugation treatment to obtain cell sediment;

[0096] (3) Suspension treatment: add the cell sediment after enzymatic hydrolysis treatment in step (2) to the suspension for treatment;

[0097] (4) Cryopreservation treatment: Suspend the hematopoietic stem cells treated in step (3), centrifuge and take out the cell pellet again, add stabilizing solution, and place in a -80°C refrigerator.

[0098] In the method for cryopreserving hematopoietic stem cells described above,

[0099] The hematopoietic stem cells described in step (1) were purchased from Beijing Zeping Techn...

Embodiment 3

[0127] The hematopoietic stem cell cryopreservation method of the present embodiment comprises the following steps:

[0128] (1) Pretreatment: Prepare hematopoietic stem cells and suspend them with normal saline;

[0129] (2) Enzymatic hydrolysis treatment: transfer the hematopoietic stem cells pretreated in step (1) to an enzymatic hydrolysis tank, perform enzymatic hydrolysis treatment, and then perform centrifugation treatment to obtain cell sediment;

[0130] (3) Suspension treatment: add the cell sediment after enzymatic hydrolysis treatment in step (2) to the suspension for treatment;

[0131] (4) Cryopreservation treatment: Suspend the hematopoietic stem cells treated in step (3), centrifuge and take out the cell pellet again, add stabilizing solution, and place in a -80°C refrigerator.

[0132] In the method for cryopreserving hematopoietic stem cells described above,

[0133] The hematopoietic stem cells described in step (1) were purchased from Beijing Zeping Techn...

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Abstract

The invention discloses a hematopoietic stem cell cryopreservation method, and belongs to the technical field of medical manufacturing. The method comprises the following steps: (1) pretreatment: preparing hematopoietic stem cells, and performing suspension treatment by using normal saline; (2) enzymolysis treatment: transferring the hematopoietic stem cells pretreated in the step (1) into an enzymolysis tank, performing enzymolysis treatment, then performing centrifugal treatment, and taking a cell precipitate; (3) suspension treatment: adding the cell precipitate subjected to enzymolysis treatment in the step (2) into a suspension for treatment; and (4) cryopreservation treatment: centrifuging the hematopoietic stem cells subjected to suspension treatment in the step (3), taking out cell precipitates again, adding a stabilizing solution, and performing placing in a refrigerator at the temperature of minus 80 DEG C. According to the hematopoietic stem cell cryopreservation method, the cryopreservation use effect of the hematopoietic stem cells can be effectively improved.

Description

technical field [0001] The invention belongs to the technical field of medical manufacture, and in particular relates to a method for cryopreserving hematopoietic stem cells. Background technique [0002] Hematopoietic stem cells are adult stem cells in the blood system. They are a heterogeneous group with the ability of long-term self-renewal and the potential to differentiate into various mature blood cells. It is a type of adult stem cell with the longest and most in-depth research history, and it has important guiding significance for the study of various stem cells, including tumor stem cells. Mature cells in the blood system have a very short lifespan. Therefore, in a person's life, hematopoietic stem cells need to replenish various mature cell components of the blood system in a timely manner according to the physiological needs of the body. At the same time, under stress conditions such as injury and inflammation, hematopoietic stem cells also play a role in regulat...

Claims

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Application Information

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IPC IPC(8): C12N5/0789A01N1/02
CPCC12N5/0647A01N1/0221A01N1/0226C12N2509/00
Inventor 不公告发明人
Owner 合肥滴碧云生物科技有限公司