Unlock instant, AI-driven research and patent intelligence for your innovation.

Pluripotent stem cell cryopreservation liquid, application thereof and cryopreservation method

A technology of pluripotent stem cells and cryopreservation solution, applied in the field of pluripotent stem cell cryopreservation solution, can solve the problems of unsuitable regenerative medicine products, infection, etc., and achieve the effects of expanding clinical application prospects, convenient use, and simple configuration.

Active Publication Date: 2021-03-26
IREGENE THERAPEUTICS LTD
View PDF19 Cites 3 Cited by
  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Although many scientists use a variety of media containing serum components to reduce the concentration of serum components, this method of protecting stem cell activity through a large amount of animal serum is not suitable for the development of regenerative medicine products. Serum components will carry a variety of animal-derived ingredients, or viruses, that can cause serious infections and complications

Method used

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
View more

Image

Smart Image Click on the blue labels to locate them in the text.
Viewing Examples
Smart Image
  • Pluripotent stem cell cryopreservation liquid, application thereof and cryopreservation method
  • Pluripotent stem cell cryopreservation liquid, application thereof and cryopreservation method
  • Pluripotent stem cell cryopreservation liquid, application thereof and cryopreservation method

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0037] Example 1 Preparation of Human-derived Pluripotent Stem Cell Liquid Freezing Solution iRePlur-F

[0038] Configure iRePlur-F cell freezing medium according to the following formula, hereinafter referred to as iRePlur-F:

[0039] Duchenne's modified Eagle's medium (DMEM medium), vitamin B12 (1.2μmol / L), vitamin C (L-ascorbic acid, 64mg / L), progesterone (Progesterone, 6.3ng / ml), putrescine (Putrescine , 23ug / ml), sodium chloride (Sodium Chloride, 0.5g / L), sodium selenite (13.6ug / L), IGF (22ug / ml), Optiferrin (0.5-10μmol / L), plant source recombinant human Basic growth factor (OsrbFGF, 50ng / mL), TGF-b (1.74ng / ml), 2.5%-10% DMSO, Y-276322HCl (10nmol / L-10μmol / L).

[0040] Among them, the control experiment used DMSO Free GMP grade (ZENOAQ) freezing solution, hereinafter referred to as CELLBANKER.

Embodiment 2

[0041] Example 2 Cryopreservation of human-derived pluripotent stem cells

[0042] Human pluripotent stem cells include embryogenic pluripotent stem cells, such as H9 cell lines and human induced pluripotent stem cells. Among them, human induced pluripotent stem cells are obtained from CD34+ cell reprogramming according to "reprogramming medium and culture method of reprogramming induced pluripotent stem cells" (ZL201910050800.7).

[0043] Human pluripotent stem cells were coated with Matrigel (STEMCELL Technologies) in T25 cell culture flasks, and placed in a 37°C incubator for more than one hour after plating. According to 1×10 6 The number of cells per flask was inoculated into T25 culture flasks for expansion and passage. Cultured in commercial TeSRTM-e8 (STEMCELL Technologies) medium at 37°C, 5% CO 2 cultivated in. When the cell growth reaches 70% coverage, use 0.05% trypsin / EDTA, incubate at 37°C for 5 minutes for digestion, centrifuge and wash the digested cells at ...

Embodiment 3

[0044] Example 3 Recovery of Human-derived Pluripotent Stem Cells

[0045] 4.1 Effect of DMSO concentration on cell viability of human pluripotent stem cells after resuscitation

[0046] Human pluripotent stem cells were coated with Matrigel (STEMCELL Technologies) in T25 cell culture flasks, and placed in a 37°C incubator for more than one hour after plating. The cells frozen using the iRePlur-F freezing solution of the present invention and the CELLBANKER freezing solution of the control group were quickly thawed in a 37°C water bath. After thawing, the cells were washed with DMEM medium to remove DMSO, and the digested cells were centrifuged and washed according to 1×10 6 Inoculate T25 cell culture plates at a density of 10 ml at 37°C, 5% CO 2 cultured in a cell culture incubator. The shape of pluripotent stem cells after recovery is as follows: figure 1 shown. in, figure 1 a. figure 1 b. figure 1 c respectively show the cell morphology of human pluripotent stem cell...

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
Login to View More

PUM

No PUM Login to View More

Abstract

The invention discloses a pluripotent stem cell cryopreservation liquid, application thereof and a cryopreservation method, and relates to the technical field of biology. The cryopreservation liquid comprises inorganic salt, vitamins, a chemical small molecule inhibitor, cytokines, DMSO and other components, is clear in components, and does not contain serum or animal-derived components. Accordingto the stem cell cryopreservation liquid, the cell cryopreservation activity is greatly improved, the cryopreservation process is simple and convenient, the cells are recovered after being cryopreserved for a long time, and various characteristics of the cells are stable. Therefore, the cryopreservation liquid disclosed by the invention greatly expands the clinical application of the human-derived pluripotent stem cells.

Description

【Technical field】 [0001] The invention relates to the field of biotechnology, in particular to a cryopreservation solution for pluripotent stem cells, its application and a cryopreservation method. 【Background technique】 [0002] Regenerative medicine refers to the emerging science of using a variety of new technical disciplines to rebuild aging or functionally lost tissues and organs, and to treat related diseases through a variety of medical methods. The important research direction of regenerative medicine is the mechanism of normal tissue characteristics and functions, the biological basis of post-traumatic repair, the regeneration mechanism of tissues and organs and the differentiation mechanism of various stem cells, so as to finally obtain effective biological treatment methods. Its research methods integrate a variety of means, including the principles and methods of life science, material science, clinical medicine and other disciplines, and comprehensively solve cl...

Claims

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
Login to View More

Application Information

Patent Timeline
no application Login to View More
Patent Type & Authority Applications(China)
IPC IPC(8): A01N1/02
CPCA01N1/0221
Inventor 魏君蔡萌曹培周佳候梦莹
Owner IREGENE THERAPEUTICS LTD