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A cryopreservation solution for long-term preservation of adult stem cells and preparation method thereof

A technology for long-term preservation of adult stem cells, applied in the field of stem cell preservation, cryopreservation solution for long-term preservation of adult stem cells and its preparation, can solve the problems of stem cell oxidative damage, reduction of differentiation potential, cell damage, etc. Good effect of death and freezing

Inactive Publication Date: 2016-09-21
河北佑仁生物科技有限公司 +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, dimethyl sulfoxide can damage the cells, and it is easy to cause oxidative damage and reduce the differentiation potential of stem cells during the resuscitation process.

Method used

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  • A cryopreservation solution for long-term preservation of adult stem cells and preparation method thereof
  • A cryopreservation solution for long-term preservation of adult stem cells and preparation method thereof
  • A cryopreservation solution for long-term preservation of adult stem cells and preparation method thereof

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Experimental program
Comparison scheme
Effect test

Embodiment 1

[0013] Embodiment 1: the preparation of cryopreservation liquid

[0014] Formula: containing DMEM medium, dimethyl sulfoxide, Knock-out serum (Gibco, 10828028, the same below) and compound (I); in every 100ml of frozen storage solution, the dimethyl sulfoxide accounts for The volume percentage content is 9%, the volume percentage content of the Knock-out serum accounting for the frozen storage solution is 30%, and the balance is DMEM medium; the concentration of the compound (I) in the frozen storage solution is 15ng / ml.

[0015] Preparation method: Take the DMEM medium, dimethyl sulfoxide, Knock-out serum and compound (I) of the formula; first dissolve the compound (I) in dimethyl sulfoxide, then mix the DMEM medium, Knock-out out serum and dimethyl sulfoxide dissolved in compound (I) were mixed evenly.

[0016] Cell cryopreservation: the adult stem cells were centrifuged at 1000rpm for 10min, then the supernatant was discarded, resuspended in the cryopreservation solution...

Embodiment 2

[0019] Embodiment 2: the preparation of cryopreservation liquid

[0020] Formula: containing DMEM medium, dimethyl sulfoxide, Knock-out serum and compound (I); in every 100ml of frozen storage solution, the volume percentage of the dimethyl sulfoxide in the frozen storage solution is 8%, The Knock-out serum accounts for 20% by volume of the cryopreservation solution, and the balance is DMEM medium; the concentration of the compound (I) in the cryopreservation solution is 10 ng / ml.

[0021] Preparation method: Take the DMEM medium, dimethyl sulfoxide, Knock-out serum and compound (I) of the formula; first dissolve the compound (I) in dimethyl sulfoxide, then mix the DMEM medium, Knock-out out serum and dimethyl sulfoxide dissolved in compound (I) were mixed evenly.

[0022] Cell cryopreservation: the adult stem cells were centrifuged at 1000rpm for 10min, then the supernatant was discarded, resuspended in the cryopreservation solution containing compound (I), and put into a st...

Embodiment 3

[0025] Embodiment 3: the preparation of cryopreservation liquid

[0026] Formula: containing DMEM medium, dimethyl sulfoxide, Knock-out serum and compound (I); in every 100ml of frozen storage solution, the volume percentage of the dimethyl sulfoxide in the frozen storage solution is 10%, The Knock-out serum accounts for 40% by volume of the cryopreservation solution, and the balance is DMEM medium; the concentration of the compound (I) in the cryopreservation solution is 20 ng / ml.

[0027] Preparation method: Take the DMEM medium, dimethyl sulfoxide, Knock-out serum and compound (I) of the formula; first dissolve the compound (I) in dimethyl sulfoxide, then mix the DMEM medium, Knock-out out serum and dimethyl sulfoxide dissolved in compound (I) were mixed evenly.

[0028] Cell cryopreservation: the adult stem cells were centrifuged at 1000rpm for 10min, then the supernatant was discarded, resuspended in the cryopreservation solution containing compound (I), and put into a s...

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Abstract

Disclosed are a cryopreservation media for long-term storage of an adult stem cell and a preparation method thereof. The cryopreservation media comprises a DMEM culture media, dimethyl sulfoxide, a knock-out serum, and compound (I). For every 100 mL of the cryopreservation media, the dimethyl sulfoxide has a volume percent of 8-10%, the knock-out serum has a volume percent of 20-40%, and the remainder being the DMEM culture media. The concentration of the compound (I) in the cryopreservation media is 10-20 ng / mL. The cryopreservation media further comprises a natural product with a novel structure as an additive. A standard fetal bovine serum is replaced with the knock-out serum. The invention improves anti-apoptosis and anti-oxidation capacities of an adult stem cell after reviving, removes impacts caused by an exogenous animal serum, and provides excellent cryopreservation performance.

Description

technical field [0001] The invention belongs to the field of cell engineering and relates to the preservation of stem cells, in particular to a cryopreservation solution for long-term preservation of adult stem cells and a preparation method thereof. Background technique [0002] Stem cells are a kind of primitive cells that can form and maintain various functions, construct tissues, organs or organisms. During individual growth and development, there are two types of stem cells: embryonic stem cells and adult stem cells. Embryonic stem cells are cells that can form living individuals and be passed down during early embryonic development. From the offspring cells formed after fertilized egg division to the inner cell group in the blastocyst, they all have the characteristics of embryonic stem cells and can be separated to form embryonic stem cells. Adult stem cells are located in mature tissues and organs, and they can differentiate to form functional cells of correspondin...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): A01N1/02
CPCA01N1/02
Inventor 寇佳琳李红臣黄前荣张永强
Owner 河北佑仁生物科技有限公司
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