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Kit for animal cell digestive inoculation and application thereof

A technology of animal cells and kits, applied in animal cells, tissue cells/virus culture devices, biochemical equipment and methods, etc., can solve the problems of affecting the activity of seed cells, unfavorable uniform distribution of cells, and difficult to seed cells, etc. Achieve low cost, wide range of applications, high yield and high activity

Inactive Publication Date: 2007-01-17
EAST CHINA UNIV OF SCI & TECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0012] The main deficiency of the above digestion method is: the digestion time is too long, thus affecting the activity of seed cells
On the other hand, it is also difficult to obtain monodisperse seed cells. Many cells exist in the form of cell clusters, which is not conducive to the uniform distribution of cells on the carrier in the lower reactor culture, and affects the growth of cells.

Method used

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  • Kit for animal cell digestive inoculation and application thereof
  • Kit for animal cell digestive inoculation and application thereof
  • Kit for animal cell digestive inoculation and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1～4

[0033]The BHK, CHSE, CHO and Vero seed cells cultured by microcarriers were digested with the kit and digestion method of the present invention. Realize inoculation from 1 liter reactor to 5 liter reactor. The 1-liter reactor is for microcarrier culture with a working volume of 0.7 liters, and the 5-liter reactor is for fixed-bed or microcarrier culture with a working volume of 3.5 liters. The EDTA in the washing solution, the trypsin concentration in the digestion solution, and the collagenase concentration are shown in Table 1.

[0034] The seed suspension was pressed from the 1 L reactor into the digestion reactor, and the culture supernatant was drained from the bottom of the digestion reactor. The washing solution preheated to 37°C was introduced from the bottom and discharged from the bottom by pressure. The washing time and stirring intensity are shown in Table 1. The digestion liquid preheated to 37°C was introduced from the bottom of the digestion reactor, and the d...

Embodiment 5

[0039] The kit of the present invention can also be used for inoculation and digestion of square bottle culture. The volume of the square bottle is 75ml, the liquid volume is 10ml, and the cultured cells are Vero cells. When the cell monolayer is full, pour out the medium, add 5ml of washing solution, shake gently twice, pour out the washing solution, and add digestion After standing for 3 minutes, pour out the digestion solution, add 10ml of medium, blow the cells off the growth surface of the bottle with a pipette, and transfer them to a culture bottle of the same size at a ratio of 1:10. After 4 days, the cell monolayer Overgrown. The digestion conditions are shown in Table 3, and the digestion results are shown in Table 4.

[0040] cell

[0041] Seed bottle cell density

[0042] Compared with conventional digestion techniques, the dispersion of cells is significantly improved, which is conducive to the adherence and rapid growth of cells after passage....

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Abstract

The present invention relates to a kit for animal cell culture and extension inoculation and its application.

Description

technical field [0001] The invention relates to a kit for expanding animal cell culture and its application. Background technique [0002] Most of the animal cell lines used for the production of recombinant proteins, recombinant viruses for gene therapy and viral vaccines have anchorage-dependent properties, and these cells must grow on the surface of the carrier. In the process of industrial animal cell culture, a series of step-by-step reactors are required to cultivate seed cells before reaching the final production scale. Inoculation is the key to the expansion of adherent cells. Therefore, we have invented a digestion reactor for animal cell culture expansion inoculation, and the present invention is based on the digestion process of the digestion reactor. [0003] There are three main methods of inoculation reported in the literature: [0004] 1. Digestion and inoculation. The main process is to use a large number of square bottles or roller...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12M3/00C12N5/06C12N5/07
Inventor 孙祥明张元兴
Owner EAST CHINA UNIV OF SCI & TECH