Blood serum-free nano material cryopreserving agent, preparation method and application method thereof and cryopreserving solution

Abstract

The invention provides a blood serum-free nano material cryopreserving agent, a preparation method and an application method thereof. The cryopreserving agent comprises DMSO, a blood serum substitution and graphene oxide. The cryopreserving agent is free of any blood serum components and graphene oxide is compounded at the same time to adjust the character of the solution, so that the solution ismore favorable for storing cells, and the cryopreserving effect is also more excellent. Compared with a conventional cryopreserving agent, the survival rate of resurgent cells is improved obviously, so that the cryopreserving agent is worth being widely popularized and applied.

Description

technical field

[0001] The invention relates to the field of cell cryopreservation, in particular to a serum-free nanomaterial cryopreservation agent, a preparation method, a use method thereof, and a cryopreservation solution. Background technique

[0002] Cell cryopreservation is a technique in which cells are placed in a low temperature environment to reduce cell metabolism for long-term storage. Cell cryopreservation is one of the main methods of cell preservation, which plays a role in cell preservation.

[0003] Cell cryopreservation is one of the main methods of cell preservation. The use of cryopreservation technology can temporarily remove cells from the growth state and preserve their cell characteristics, so that cells can be revived for experiments when needed.

[0004] In the current prior art, the main components in the cell cryopreservation solution are 10% dimethylsulfoxide (DMSO), 20%-90% fetal bovine serum (FBS), and the remaining components are incomplete...

Examples

Embodiment 1

[0044] 1. Main ingredients: cell suspension and cell cryopreservation agent.

[0045] The cell suspension consists of 100% serum-free medium.

[0046] The cell cryopreservation agent is composed of 10% DMSO by volume and 90% serum substitute, adding 5ug / ml graphene oxide aqueous solution (water is the solvent, the particle size is 200nm, and the concentration is 1ug / ml)

[0047] Serum replacement model (Gibco KNOCKOUT TM Serum Replacement 10828-028).

[0048] 2. Preparation method:

[0049] Under sterile conditions, take the serum substitute and put it in a centrifuge tube, add DMSO and mix evenly, the volume ratio is serum substitute:DMSO=9:1. After taking out 5 μl of the mixture, add 5 ug / ml graphene oxide aqueous solution, mix well, and store at 4°C.

[0050] 3. How to use:

[0051] 1) In the sterile environment of the cell ultra-clean bench, add cell suspension to the 1.5ml EP tube to evenly suspend the cells, and the final concentration of cells is 1×10 6 cells / ml, ...

Embodiment 2

[0054] 1. Main ingredients: cell suspension and cell cryopreservation agent.

[0055] The cell suspension consists of 100% serum-free medium.

[0056] The cell cryopreservation agent is composed of 5% DMSO by volume and 95% serum substitute, adding 1ug / ml graphene oxide aqueous solution (water is the solvent, the particle size is 20nm, and the concentration is 5ug / ml)

[0057] Serum replacement model (Gibco KNOCKOUT TM Serum Replacement 10828-028).

[0058] 2. Preparation method:

[0059] Under sterile conditions, take the serum substitute and put it in a centrifuge tube, add DMSO and mix evenly, the volume ratio is serum substitute:DMSO=15:1. After taking out 5 μl of the mixture, add 1 ug / ml graphene oxide aqueous solution, mix well, and store at 4°C.

[0060] 3. How to use:

[0061] 1) In the sterile environment of the cell ultra-clean bench, add cell suspension to the 1.5ml EP tube to evenly suspend the cells, and the final concentration of cells is 6×10 6 cells / ml, t...

Embodiment 3

[0064] 1. Main ingredients: cell suspension and cell cryopreservation agent.

[0065] The cell suspension consists of 100% serum-free medium.

[0066] The cell cryopreservation agent is composed of 9% DMSO by volume and 91% serum substitute, adding 2ug / ml graphene oxide aqueous solution (water is the solvent, the particle size is 100nm, and the concentration is 3ug / ml)

[0067] Serum replacement model (Gibco KNOCKOUT TM Serum Replacement 10828-028).

[0068] 2. Preparation method:

[0069] Under sterile conditions, take the serum substitute and put it in a centrifuge tube, add DMSO and mix evenly, the volume ratio is serum substitute:DMSO=1:10. After taking out 5 μl of the mixture, add 2ug / ml graphene oxide aqueous solution, mix well, and store at 4°C.

[0070] 3. How to use:

[0071] 1) In the sterile environment of the cell ultra-clean bench, add cell suspension to the 1.5ml EP tube to evenly suspend the cells, and the final concentration of cells is 5×10 6 cells / ml, t...

technical field

[0001] The invention relates to the field of cell cryopreservation, in particular to a serum-free nanomaterial cryopreservation agent, a preparation method, a use method thereof, and a cryopreservation solution. Background technique

[0002] Cell cryopreservation is a technique in which cells are placed in a low temperature environment to reduce cell metabolism for long-term storage. Cell cryopreservation is one of the main methods of cell preservation, which plays a role in cell preservation.

[0003] Cell cryopreservation is one of the main methods of cell preservation. The use of cryopreservation technology can temporarily remove cells from the growth state and preserve their cell characteristics, so that cells can be revived for experiments when needed.

[0004] In the current prior art, the main components in the cell cryopreservation solution are 10% dimethylsulfoxide (DMSO), 20%-90% fetal bovine serum (FBS), and the remaining components are incomplete...

Examples

Embodiment 1

[0044] 1. Main ingredients: cell suspension and cell cryopreservation agent.

[0045] The cell suspension consists of 100% serum-free medium.

[0046] The cell cryopreservation agent is composed of 10% DMSO by volume and 90% serum substitute, adding 5ug / ml graphene oxide aqueous solution (water is the solvent, the particle size is 200nm, and the concentration is 1ug / ml)

[0047] Serum replacement model (Gibco KNOCKOUT TM Serum Replacement 10828-028).

[0048] 2. Preparation method:

[0049] Under sterile conditions, take the serum substitute and put it in a centrifuge tube, add DMSO and mix evenly, the volume ratio is serum substitute:DMSO=9:1. After taking out 5 μl of the mixture, add 5 ug / ml graphene oxide aqueous solution, mix well, and store at 4°C.

[0050] 3. How to use:

[0051] 1) In the sterile environment of the cell ultra-clean bench, add cell suspension to the 1.5ml EP tube to evenly suspend the cells, and the final concentration of cells is 1×10 6 cells / ml, ...

Embodiment 2

[0054] 1. Main ingredients: cell suspension and cell cryopreservation agent.

[0055] The cell suspension consists of 100% serum-free medium.

[0056] The cell cryopreservation agent is composed of 5% DMSO by volume and 95% serum substitute, adding 1ug / ml graphene oxide aqueous solution (water is the solvent, the particle size is 20nm, and the concentration is 5ug / ml)

[0057] Serum replacement model (Gibco KNOCKOUT TM Serum Replacement 10828-028).

[0058] 2. Preparation method:

[0059] Under sterile conditions, take the serum substitute and put it in a centrifuge tube, add DMSO and mix evenly, the volume ratio is serum substitute:DMSO=15:1. After taking out 5 μl of the mixture, add 1 ug / ml graphene oxide aqueous solution, mix well, and store at 4°C.

[0060] 3. How to use:

[0061] 1) In the sterile environment of the cell ultra-clean bench, add cell suspension to the 1.5ml EP tube to evenly suspend the cells, and the final concentration of cells is 6×10 6 cells / ml, t...

Embodiment 3

[0064] 1. Main ingredients: cell suspension and cell cryopreservation agent.

[0065] The cell suspension consists of 100% serum-free medium.

[0066] The cell cryopreservation agent is composed of 9% DMSO by volume and 91% serum substitute, adding 2ug / ml graphene oxide aqueous solution (water is the solvent, the particle size is 100nm, and the concentration is 3ug / ml)

[0067] Serum replacement model (Gibco KNOCKOUT TM Serum Replacement 10828-028).

[0068] 2. Preparation method:

[0069] Under sterile conditions, take the serum substitute and put it in a centrifuge tube, add DMSO and mix evenly, the volume ratio is serum substitute:DMSO=1:10. After taking out 5 μl of the mixture, add 2ug / ml graphene oxide aqueous solution, mix well, and store at 4°C.

[0070] 3. How to use:

[0071] 1) In the sterile environment of the cell ultra-clean bench, add cell suspension to the 1.5ml EP tube to evenly suspend the cells, and the final concentration of cells is 5×10 6 cells / ml, t...

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