Blood serum-free nano material cryopreserving agent, preparation method and application method thereof and cryopreserving solution
A technology of nanomaterials and cryopreservatives, which is applied in applications, preservation of human or animal bodies, animal husbandry, etc. It can solve problems such as low rewarming efficiency, large cell damage, and complex components, and achieves mild operating conditions and no three wastes Produce and operate simple effects
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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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