A kind of fetal bovine serum substitute and its preparation method and application

A technology of fetal bovine serum and substitutes, applied in biochemical equipment and methods, microorganisms, tissue culture, etc., can solve the problems of high production and use costs, and achieve good growth status and good inter-batch stability

Active Publication Date: 2018-12-21
GUANGZHOU RUITE BIOTECH CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Because the fetal cow has not been in contact with the outside world, the serum contains the least harmful components such as antibodies and complements, and the quality is the highest. Correspondingly, its production and use costs are also extremely high

Method used

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  • A kind of fetal bovine serum substitute and its preparation method and application
  • A kind of fetal bovine serum substitute and its preparation method and application
  • A kind of fetal bovine serum substitute and its preparation method and application

Examples

Experimental program
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Effect test

Embodiment 1

[0049] A kind of fetal calf serum substitute, its preparation method is as follows:

[0050] 1) Choose robust adult cows, take 5 liters of blood as a unit, add anticoagulant (4% trisodium citrate dihydrate) to 500ml; put it in a refrigerator at 4°C overnight, and take yellow plasma under sterile conditions , centrifuge at 3000rpm at 4°C for 10min and take the supernatant;

[0051] 2) Add CaCl with a final concentration of 20mM to the supernatant 2 , 37°C water bath for 2h;

[0052] 3) After standing at -20°C for 12 hours, place at room temperature to melt, remove the protein precipitate and collect the supernatant;

[0053] 4) Add NaCl with a final concentration of 4.79M to the supernatant, and precipitate at 4°C for 12 hours;

[0054] 5) Centrifuge at 5000rpm for 15min and collect the supernatant;

[0055] 6) Desalting, adjusting the NaCl concentration to 0.7%;

[0056] 7) Add ions necessary for cell growth, and the final concentration is:

[0057]

[0058]

[005...

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Abstract

A fetal bovine serum substitute, a preparation method therefor, and an application thereof in a culture medium. The fetal bovine serum substitute is prepared by adding nutrient elements to a protein-depleted bovine serum. The nutrients consist of a specific ratio of CaCl2•2H2O, CuSO4•5H2O, 1% ferrous citrate, hydrocortisone, FeSO4•7H2O, KCl, MgCl2•6H2O, MgSO4•7H2O, NaCl, NaH2PO4•2H2O, Na2HPO4•12H2O, glucose, sodium pyruvate, a GABA stock solution, a LiCl stock solution, a piperidic acid concentrate, a Stock A stock solution, and a Na2SeO3 stock solution.

Description

technical field [0001] The invention relates to a biological product, in particular to a fetal bovine serum substitute and its preparation method and application. Background technique [0002] Animal serum is the most used natural medium in cell culture. It is rich in nutrients necessary for cell growth and is often used for in vitro culture of animal cells. [0003] Animal serum mainly includes fetal bovine serum, calf serum, adult bovine serum, horse serum, goat serum, chicken serum, rabbit serum, etc. Among them, fetal bovine serum is the most commonly used. Animal serum provides growth factors, hormones, transfer proteins, factors for attachment and spreading on the culture substrate, protease inhibitors and other nutrients required for cell proliferation in cell culture. The advantage of using serum is that serum contains most of the factors that promote cell proliferation and maintenance, and it is almost a universal growth supplement, suitable for cell culture of ani...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N5/00
CPCC12N5/0037
Inventor 李长路
Owner GUANGZHOU RUITE BIOTECH CO LTD
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