SU162294A1 - - Google Patents

Description

It is known to use nutrient protein-free supporting media for animal cells grown in vitro, containing protein hydrolyzate, saline and vitamins. These media provide a satisfactory cell survival time only in the case of regular (twice weekly) medium renewal.

In order to increase the cell survival time, nucleated sodium is introduced into the protein-free nutrient medium. For the preparation of the proposed nutrient-free protein supportive medium, the salt solution (89.88-89.84%), the protein hydrolyzate (10.0%), the solution of vitamins (0.05% folic acid solution), and sodium nucleic acid (0 , 02- 0.06%).

As a saline solution, any of the cells commonly used for culturing is used, for example, Hanks or Earl saline solution. Sterile liquid protein hydrolysates used for parenteral nutrition of patients can be used as a protein hydrolyzate. The use of a solution of lactalbumin hydrolyzate and other protein hydrolysates is also not excluded, as long as they contain all the amino acids necessary for cell growth.

Example 1. To prepare a solution of vitamins, 2 ml of concentrated hydrochloric acid and 10 ml of riboflavin are added to 10 ml of desalted water. To completely dissolve the riboflavin, the mixture is heated in a boiling water bath. The cooled solution is made up to 100 ml with desalted water and 100 mg of pyridoxia (or pyridoxal), calcium pantothenate (or pantothenic acid), thiamine, pyacin and choline chloride and 200 mesoinzite are added. After complete dissolution of all components of the solution, its volume is brought to 198 ml with chemically pure or distilled glycerol and 2 ml of a 1.7% solution of nystatin in a 70% standard are added. Store the solution in an opaque dish at 15 ° C. Shelf life 6 months. The solution can be consumed no earlier than 5 days after its manufacture.

Example 2. To prepare a folic acid solution, 10 ml of a 7.5% sodium bicarbonate solution are added to 90 ml of desalted water and 100 mg of folic acid is added, after which 80 mg of tetracycline hydrochloride is added. The volume of the solution is made up to 198 ml with chemically pure glycerol and 2 ml of a 1.7% solution of nystatin in a 70% standard are added. The storage conditions, shelf life and shelf life before consumption are the same as for vitamins.

Example To prepare a solution of nucleic acid sodium, 2000 mg of salt are added to 10 ml of desalted water, after dissolving which the volume is brought to 18 ml with chemically pure glycerin, and 2 ml of nystatin solution is added. Storage conditions, term

The shelf life and durability before use are the same as for the solution of vitamins.

When testing the cultural properties of the proposed supportive environment using as a test object of permanent cells of SOC, permanent cells of human amnion and primary trypsinized cells of a human embryo, the periods of experience were 14-18, 14-17 and 16-21 days, respectively. In case of absence in the described support, her environment and the acid-acidic patri cell survived no more than 7 days.

The subject invention 1i

A nutrient-free protein-free support for animal cells grown in vitro, consisting of protein hydrolyzate, saline, vitamins, characterized in that nucleic sodium is introduced into its composition in order to prolong the cell survival time.