Cell cryopreservation composition and cryopreservation method

Inactive Publication Date: 2020-02-13
SARAYA CO LTD +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Benefits of technology

[0015]Addition of SL can reduce freezing damage to cells. As a result, a certain level or hi

Problems solved by technology

However, it is known that, in the process of freezing cells, water in and out of the cells turns into ice crystals and the ice crystals damage the cells (Non-Patent Document 1).
However, a cryopreservation solution containing DMSO as a cryoprotectant does not have satisfactory preservation efficiency, and it cannot necessarily be said that this cryopreservation solution sufficiently inhibits ice crystal formation.
However, it cannot be said that these cryop

Method used

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  • Cell cryopreservation composition and cryopreservation method
  • Cell cryopreservation composition and cryopreservation method
  • Cell cryopreservation composition and cryopreservation method

Examples

Experimental program
Comparison scheme
Effect test

example 14

[0047 had a higher proliferation rate than Comparative Example 7.

[0048]Human normal fibroblasts were sown on a 96-well plate at 2.0×104 cells / ml and cultured for 6 or 72 hours. After culturing, absorbance was measured with Cell Counting Kit-8 (DOJINDO LABORATORIES) (absorbance before freezing). The remaining cells were suspended at 4.0×105 cells / ml in DMEM containing fetal bovine serum. Each of the compositions shown in Table 9 and the cell suspension were mixed at a volume ratio of 1:1 in CRYOGENIC VIAL (Sansyo Co., Ltd.). Each of the resultant cell suspensions was placed in a freezing container, BICELL (Nihon Freezer Co., Ltd.), and cooled at −80° C. After overnight storage, each cell suspension was rapidly thawed at 37° C., 100 μl of each cell suspension was sown on a 96-well plate, and the cells were cultured for 6 or 72 hours. Absorbance was then measured with Cell Counting Kit-8 (DOJINDO LABORATORIES) (absorbance after cryopreservation).

[0049]The viability and the proliferatio...

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Abstract

This composition can be easily added when cryopreserving cells and improves cell viability. 1 volume % of the composition containing 0.01 wt % to 20 wt % of a sophorose lipid is added just before or up to 6 hours before cryopreservation and cells are stored. This composition can improve cryopreservation cell viability as it reduces freezing damage to cells. With this composition, cells can be stored without using DMSO and blood serum. This is a simple and inexpensive storage method as it does not require fine control of the freezing rate during cell cryopreservation.

Description

TECHNICAL FIELD[0001]The present invention relates to cryopreservation compositions that are used to cryopreserve cells, and cryopreservation methods using the same.BACKGROUND ART[0002]Cell cryopreservation is widely used as an essential technique to prevent cell degeneration due to passaging, prevent contamination with bacteria associated with passaging, transport cells, etc. However, it is known that, in the process of freezing cells, water in and out of the cells turns into ice crystals and the ice crystals damage the cells (Non-Patent Document 1). It I therefore desired to protect cells from damage during freezing and thawing and to cryopreserve cells while maintaining their properties.[0003]In common cryopreservation, cells are typically suspended in a culture solution containing bovine serum etc. with a cryoprotectant for protecting the cells from damage from ice crystals, and the cell suspension thus obtained is placed in a cryotube etc., cooled, and eventually cryopreserved ...

Claims

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Application Information

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IPC IPC(8): A01N1/02C12N1/04C12N5/0775
CPCA01N1/0221C12N5/0665C12N1/04C12N5/0663C12N5/0018C12N5/0662C12N2500/30C12N2500/35C12N2500/62
Inventor NOGAMI, ASUKATATSUMI, MOTOKIISHII, NANASERYU, MIZUYUKIHIRATA, YOSHIHIKOSAWA, YOSHIKIMIYAGAWA, SHIGERUSAITO, ATSUHIROOHKAWARA, HIROTATSU
Owner SARAYA CO LTD
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