CS258253B1 - Method of increasing mitotic index - Google Patents

Abstract

Using compounds whose cation has the general formula N C H ” COOH where R", R" is alkyl, with 1 to 6 carbon atoms, an increase in the mitotic index to more than three times the usual value in tissue, cell or protoplast cultures is achieved. The method of increasing the mitotic index and partial synchronization of the cell population in the first half of the culture cycle is significant for the biochemical regulation of biotechnological processes in the pharmaceutical, food and feed industries.

Description

The invention relates to the biochemical regulation of biotechnological processes.

Biotechnological processes using tissue and / or cell and / or protoplast cultures are still encountering difficulties associated with insufficiently synchronous division of the cell population.

As follows from the following example, the use of a cation of the general formula

R.

RCH = N ⁺ H COOH where

R ', R is an alkyl of 1 to 6 carbon atoms, is based on the finding of a new property, namely the ability to increase the value of the mitotic index in the first half of the cultivation cycle. It can be reasonably assumed that a short-term but significant increase in the mitotic index results in a synchronizing effect in the cell population. The present invention provides a method for increasing the mitotic index and / or partial synchronization of nuclear and / or cell divisions in tissue and / or cell and / or protoplast cultures, characterized in that a compound of the formula

COOH where

R ', R is alkyl of 1 to 6 carbon atoms, at a final concentration of 10 ^-13 to 10 ^-4 .

This also achieves a significant increase in the number of cells that is significant before it reaches the middle of the culture cycle. This is an important step to shorten the biotechnology production cycle, since the desired cell concentration per unit volume can be achieved more quickly.

Example

To a standard suspension culture solution optimized for Papaver somniferum (opium alkaloid producer) cells, aqueous solutions freshly prepared from fresh para- (N-dimethylaminobenzylidene) -para-aminobenzoic acid sulfate crystals were added prior to inoculation.

CH = N ⁺ H

HSOj

to achieve a concentration of said compound of 0.5 mg per liter of nutrient solution. The initial cell concentration of the inoculation was 70,000 per ml suspension. The culture cycle lasted 15 days. The table below shows the mitotic index as a function of time from the start of the culture cycle

Table

Number of days since the beginning of cultivation Mitotic index Control With the addition of 0.5 mg of the above compound per 1 liter of suspension 0 0.20 0.20 3 1.13 1.24 5 1.84 6.21 8 3.44 3.48 15 Dec 0.17 0.26

Thus, according to the invention, a mitotic index of 6.21 4 can be achieved while a mitotic index of at most 3.44 4 has been achieved in the control. On the fifth day of cultivation according to the invention a mitotic index value of 6.21 4 is induced only 1.84 4; our invention thus allows a 3.4-fold increase. It is also important that the maximum mitotic index value in the control variant is only reached on the eighth day of the culture cycle, while the method according to the invention ensures that the maximum value is reached on the fifth day, so that the partial synchronization effect occurs three days earlier.

Claims (1)

Method for increasing the mitotic index and / or partial synchronization of cell divisions in tissue, and / or cell and / or protoplast cultures in the first half of a culture cycle, characterized in that a compound whose cation of general formula ^CH = ^{N + H COOH is} added to the culture medium wherein R ', R is alkyl of 1 to 6 carbon atoms, at a final concentration of 10 ^-12 to 10 ~ ^4th