Refrigerated preservation solution for stem cells

A technology of stem cells and preservation solution, applied in the field of preservation solution, which can solve the problems of low survival rate, slowness, and cold storage without pluripotent cells, and achieve the effect of maintaining proliferation ability and high proliferation ability

Pending Publication Date: 2022-06-03
BIOVERDE
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The cryopreservation of human iPS cells and ES cells has many problems, such as the slow cryopreservation method, the survival rate after storage is low, or DMSO, which is a freeze-resistant material used for cryopreservation, has cytotoxicity and differentiation induction Wait
There are reports that this HTSFRS can cryopreserve human skin fibroblasts or smooth muscle cells for 3 to 5 days, but there is no report on cryopreservation of pluripotent cells such as human iPS cells

Method used

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  • Refrigerated preservation solution for stem cells
  • Refrigerated preservation solution for stem cells
  • Refrigerated preservation solution for stem cells

Examples

Experimental program
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Effect test

Embodiment

[0140] 1. Preparation of cells for cryopreservation

[0141] A healthy human-derived human iPS cell line (253G1) obtained from Kyoto University's iPS cell research was maintained and cultured under feeder-free conditions. In this culture, StemFit (registered trademark) AK02N (Ajinomoto), which is a commercially available medium for clinical research (for human ES / iPS cells), was used, and the concentration of 70 to 80% confluence was reached on the seventh day after the start of culture. At the time point, stripping was performed with an enzyme (using 0.5X TrypLE (registered trademark) Select), dispersed and washed. Then, it was suspended in a new StemFit (registered trademark) AK02N medium containing 10 μM of Y27362 (Fujifilm Wako Pure Chemical Industries, Ltd.) as a ROCK inhibitor at a concentration of 0.1 μg / cm 2 Add iMatrix-511 (MATRIXOME, Inc.) as a cell culture substrate in the manner of 1.3E3 / cm 2 (1.3×10 3 / cm 2 ) concentrations were inoculated. On the second day ...

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Abstract

Provided are a cryopreservation solution and a cryopreservation method suitable for cryopreserving stem cells or embryos such as human or animal iPS cells in a non-frozen state. The refrigerated preservation liquid according to one embodiment has an ion-based molar ratio (Na + / K + ratio) of the amount of sodium ion species to the amount of potassium ion species of 0.5-1.3, contains Trolox or an analog thereof at a concentration of 1-8 mM, contains adenine or a salt or derivative thereof at a concentration of 0.1-4.2 mM, and contains all essential amino acids, or all essential amino acids other than one, two or three essential amino acids are contained, the total content of the essential amino acids is 50-200 mg / L, and the total content of the non-essential amino acids is 50-200 mg / L.

Description

technical field [0001] The present invention relates to a preservation solution for preserving cells, tissues, organoids, organs and the like in a non-frozen state. In particular, it relates to a cryopreservation solution for storage at a temperature lower than room temperature, for example, 2 to 15°C, 2 to 8°C, or 2 to 6°C. In particular, it relates to a method for suspending or immersing pluripotent stem cells (pluripotent cells) such as human iPS cells and other stem cells (stemcells), or derivatives such as organoids obtained therefrom, and primary embryos (primary embryos). refrigerated preservation solution for preservation. Background technique [0002] The simple, efficient and stable storage and supply of stem cells such as iPS and ES cells is an indispensable and important technology in the fields of basic research, clinical application research or drug development research of regenerative medicine. The cryopreservation of human iPS cells and ES cells has the fol...

Claims

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Application Information

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IPC IPC(8): A01N1/02C12N5/0735C12N5/10
CPCA01N1/0221A01N1/0226C12N5/0606C12N5/0696C12N2500/12C12N2523/00C12N2500/32C12N2500/34C12N2500/40C12N2500/38A01N1/021
Inventor 相泽明玄丞烋玄优基
Owner BIOVERDE
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