Serum-free and DMSO-free cell cryopreservation solution and preparation method thereof

A serum-free, cryopreserved liquid technology is applied in the field of cell culture to achieve the effects of high biological safety, simple preparation method and high clinical application prospect.

Inactive Publication Date: 2020-03-31
苏州君欣生物科技有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

But this is only the research prototype of serum-free cell culture medium and cell cryopreservation solution

Method used

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  • Serum-free and DMSO-free cell cryopreservation solution and preparation method thereof
  • Serum-free and DMSO-free cell cryopreservation solution and preparation method thereof
  • Serum-free and DMSO-free cell cryopreservation solution and preparation method thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0026] The serum-free DMSO-free cell cryopreservation solution has the following components: anhydrous calcium chloride 116.6 mg / L, L-leucine 59.05 mg / L, linoleic acid 0.042 mg / L, copper sulfate pentahydrate 00013mg / L, L-lysine hydrochloride 91.25mg / L, lipoic acid 0.105mg / L, ferric nitrate nonahydrate 0.05mg / L, L-methionine 17.24mg / L, phenol red 8.1mg / L, heptahydrate Ferrous sulfate 0.417mg / L, L-phenylalanine 35.48mg / L, 1,4-butylene diamine dihydrochloride 0.081mg / L, potassium chloride 311.8mg / L, L-serine 26.25mg / L , sodium pyruvate 55mg / L, magnesium chloride 28.64mg / L, threonine 53.45mg / L, vitamin H 0.0035mg / L, anhydrous magnesium sulfate 48.84mg / L, alanine 4.45mg / L, calcium pantothenate 2.24mg / L, sodium chloride 6999.5mg / L, L-asparagine 7.5mg / L, choline chloride 8.98mg / L, anhydrous sodium dihydrogen phosphate 54.35mg / L, aspartic acid 6.65mg / L, Folic acid 2.65mg / L, disodium hydrogen phosphate 71.02mg / L, L-cysteine ​​hydrochloride 17.56mg / L, I-inositol 12.6mg / L, zinc sulfate...

Embodiment 2

[0028] The serum-free DMSO-free cell cryopreservation solution has the following components: anhydrous calcium chloride 116.6 mg / L, L-leucine 59.05 mg / L, linoleic acid 0.042 mg / L, copper sulfate pentahydrate 00013mg / L, L-lysine hydrochloride 91.25mg / L, lipoic acid 0.105mg / L, ferric nitrate nonahydrate 0.05mg / L, L-methionine 17.24mg / L, phenol red 8.1mg / L, heptahydrate Ferrous sulfate 0.417mg / L, L-phenylalanine 35.48mg / L, 1,4-butylene diamine dihydrochloride 0.081mg / L, potassium chloride 311.8mg / L, L-serine 26.25mg / L , sodium pyruvate 55mg / L, magnesium chloride 28.64mg / L, threonine 53.45mg / L, vitamin H 0.0035mg / L, anhydrous magnesium sulfate 48.84mg / L, alanine 4.45mg / L, calcium pantothenate 2.24mg / L, sodium chloride 6999.5mg / L, L-asparagine 7.5mg / L, choline chloride 8.98mg / L, anhydrous sodium dihydrogen phosphate 54.35mg / L, aspartic acid 6.65mg / L, Folic acid 2.65mg / L, disodium hydrogen phosphate 71.02mg / L, L-cysteine ​​hydrochloride 17.56mg / L, I-inositol 12.6mg / L, zinc sulfate...

Embodiment 3

[0030] The serum-free DMSO-free cell cryopreservation solution has the following components: anhydrous calcium chloride 116.6 mg / L, L-leucine 59.05 mg / L, linoleic acid 0.042 mg / L, copper sulfate pentahydrate 00013mg / L, L-lysine hydrochloride 91.25mg / L, lipoic acid 0.105mg / L, ferric nitrate nonahydrate 0.05mg / L, L-methionine 17.24mg / L, phenol red 8.1mg / L, heptahydrate Ferrous sulfate 0.417mg / L, L-phenylalanine 35.48mg / L, 1,4-butylene diamine dihydrochloride 0.081mg / L, potassium chloride 311.8mg / L, L-serine 26.25mg / L , sodium pyruvate 55mg / L, magnesium chloride 28.64mg / L, threonine 53.45mg / L, vitamin H 0.0035mg / L, anhydrous magnesium sulfate 48.84mg / L, alanine 4.45mg / L, calcium pantothenate 2.24mg / L, sodium chloride 6999.5mg / L, L-asparagine 7.5mg / L, choline chloride 8.98mg / L, anhydrous sodium dihydrogen phosphate 54.35mg / L, aspartic acid 6.65mg / L, Folic acid 2.65mg / L, disodium hydrogen phosphate 71.02mg / L, L-cysteine ​​hydrochloride 17.56mg / L, I-inositol 12.6mg / L, zinc sulfate...

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Abstract

The invention discloses a serum-free and DMSO-free cell cryopreservation solution, which comprises a 16.762-16.825 g/L DMEM/F-12 basal culture medium, 32-34 mg/L cytokines, 0.2-0.8 mg/L hydroxyethyl starch, 1-3 mg/L vitamins, 0.15-0.16 g/L antibiotics, 5-6 mg/L protein polypeptide, 1-4 mg/L lipids and a 0.1-0.41 g/L intracellular and extracellular cryoprotectant. According to the technical scheme,the obtained cell cryopreservation solution does not contain serum or DMSO components, the components of the formula are determined, the cell cryopreservation solution has high biological safety andclinical application prospects, and the cell cryopreservation liquid has a cell recovery survival rate of more than 85%, can maintain good cell activity and good physiological characteristics, and issuitable for the related research fields of primary cells and stem cells.

Description

technical field [0001] The invention belongs to the technical field of cell culture, and in particular relates to a serum-free and DMSO-free cell cryopreservation solution and a preparation method thereof. Background technique [0002] Defining the media components necessary for the survival and proliferation of cultured animal cells has been a very active area of ​​research. From the initial medium formed by simple salt ions, a variety of biological factors have been added to the basal medium. Eagle and his colleagues discovered that amino acids and vitamins are required to grow mammalian cells in vitro. Later, serum was added to most synthetic media to support cell viability and proliferation in vitro. However, most of the serum is taken from exogenous fetal bovine serum. For general mammalian cell culture, as long as the growth state of the cells can be maintained, the synthetic medium can be used for cell culture. [0003] With the development of medical technology an...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A01N1/02
CPCA01N1/0221
Inventor 高宁席其良
Owner 苏州君欣生物科技有限公司
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