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Biological sample cryoprotectant and its application

A cryoprotectant, biological sample technology, applied in the field of cell preservation solution, can solve the problems of cumbersome operation, cytotoxicity, hindering cell proliferation, etc.

Active Publication Date: 2021-04-06
BEIJING TAIDONG BIOTECH CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, when these cryopreservatives are concentrated, there is usually a problem of showing toxicity to cells.
And because the above-mentioned preservative hinders the proliferation of cells, the cryopreservative must be removed from the cells in advance when culturing, resulting in cumbersome operations.

Method used

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  • Biological sample cryoprotectant and its application
  • Biological sample cryoprotectant and its application
  • Biological sample cryoprotectant and its application

Examples

Experimental program
Comparison scheme
Effect test

Embodiment approach

[0063] According to any embodiment of the present invention, the biological sample includes organs, tissues or cells.

[0064] The species source of the biological sample can be selected as animals, including humans and all livestock (such as domestic animals and pets) and wild animals and birds, which include, without limitation, cattle, horses, dairy cows, pigs, sheep, goats, Rats, mice, dogs, cats, rabbits, camels, donkeys, deer, mink, chickens, ducks, geese, turkeys, fighting cocks, etc.; the species of the biological sample can also be plants or microorganisms with cellular structures.

[0065] According to any embodiment of the present invention, the organ and tissue are selected from the group consisting of spleen, heart, liver, brain and other nervous tissues, kidney, lung, pancreas, breast, umbilical cord, skin, placenta, ovary, fallopian tube, uterus, prostate , tonsil, thymus, stomach, testis, trachea, cartilage, tendon, bone, skeletal muscle, smooth muscle, intesti...

Embodiment 1

[0071] This example is used to illustrate the application of Rbm46 as a cryoprotectant component; the biological sample to be protected is egg cells.

[0072]Unless otherwise specified, the Rbm46 used in the examples of the present application is of human (Homo sapiens) origin, and its NCBI Reference Sequence is: NP_001264100.1.

[0073] 1. Expression of Rbm46 in human oocytes

[0074] The inventor observed the distribution of Rbm46 in egg cells at different stages of meiosis and found that Rbm46 presents a point-like aggregated distribution in the cells ( figure 1 ). Several papers have previously reported that RNA-binding proteins can accumulate in cells and participate in the formation of germ cell granules. We think that this distribution of Rbm46 is very likely to participate in the formation of germ cell granules.

[0075] 2. Rbm46 is a phase transition protein

[0076] It has been previously reported that proteins involved in the formation of RNA granules in vivo ar...

Embodiment 2

[0095] Using HEK293T cells for cryopreservation test, four kinds of sample cryopreservation solutions were prepared.

[0096] The first sample freezing solution contains ethylene glycol (EG) and phosphate buffer at a concentration of 22.5% (v / v);

[0097] The second sample freezing solution contains EG [22.5% (v / v)], 5mM Rbm46 and buffer;

[0098] The third sample freezing solution contains 5mM Rbm15 and buffer;

[0099] The fourth sample freezing solution contained 5 mM Rbm47 and buffer.

[0100] When the HEK293 cells grew to 80% confluence, the cells were treated with 0.25% trypsin to form single cells, the complete medium was used to stop digestion and the cells were resuspended, centrifuged at 200g, and the supernatant was removed. Add an appropriate amount of phosphate buffer to resuspend the cells, use a hemocytometer to count the cells, and adjust the cell concentration to 1×10 6 / ml. Divide the cells into 4 centrifuge tubes, 1ml in each tube, centrifuge at 200g, an...

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Abstract

The invention relates to the field of cell preservation solution, in particular to a biological sample cryoprotectant and its application. The biological sample cryoprotectant working solution contains ≥ 5mM active fragments; the active fragments are capable of phase separation; and contain RNA binding domains and low-complexity sequence domains of polypeptides and / or proteins. The cryoprotectant for biological samples is non-toxic or low-toxic to biological samples, and can reduce the generation of ice crystals, balance the osmotic pressure inside and outside the cell membrane of biological samples, and maintain the function of biological samples after cryopreservation.

Description

technical field [0001] The invention relates to the field of cell preservation solution, in particular to a biological sample cryoprotectant and its application. Background technique [0002] Storage of cells in the frozen state (cryopreservation) is a common technique used to maintain cell viability and genetic stability in the long term. Cryopreservation is generally a process in which samples, such as cells or tissues, are preserved by cooling to a sub-zero temperature, usually 77K (= -196°C, the boiling point of liquid nitrogen). At such low temperatures, any biological activity -- including biochemical reactions that would cause cell death -- is effectively blocked. [0003] In traditional freezing techniques, samples are collected, placed in a storage solution, and then stored frozen. When samples are used, they are thawed and placed in cell culture medium. Cryopreservation protocols subject cells to various stresses and injuries (eg, damage from ice crystal formati...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): A01N1/02
CPCA01N1/0221
Inventor 李立东
Owner BEIJING TAIDONG BIOTECH CO LTD
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