Medium for culturing animal cells

Details Medium for culturing animal cells Medium for culturing animal cells

1996-08-15

1998-10-20

Degen, Nancy

Chemistry: molecular biology and microbiology

Animal cell, per se ; composition thereof; process of...

Method of culturing cells in suspension

435369, 435325, 435 703, 435 701, 435 41, C12N 500

Patent

active

058245529

DESCRIPTION:

BRIEF SUMMARY
This is the national stage of PCT/JP95/00113, filed Jan. 30, 1995.


DESCRIPTION

1. Technical Field
This invention relates to a method of culturing animal cells, and more specifically relates to a method of culturing animal cells whereby proliferation of the animal cells is enhanced and the yield of the cell itself or an useful substance produced thereby is increased, and at the same time wherein reproducibility is remarkably improved.
2. Background of the Art
Culture of animal cells is now an inevitable technique for production of various physiologically active substances, construction of drug assay systems, analysis of cell functions, etc. In culture of animal cells, is generally used a serum medium comprising a medium having added thereinto bovine serum, etc., or a so-called serum-free medium comprising a medium having added thereinto growth factors such as hormones. However, some cells do not sufficiently proliferate even by addition of a serum or addition of growth factors generally used. Therefore, it is useful to search for substances having a further growth enhancement action on these cells.
Heretofore, many substances have been reported as exhibiting a growth enhancement action on various cells, but many of them are peptides and expensive. Although it is advantageous if there are inexpensive low molecular compounds exhibiting an animal cell growth enhancement action, there are only a few reports thereon. For example, a serum-free medium or low serum medium containing p-aminobenzoic acid is disclosed as an example of media for culture of hybridomas or human or animal cell lines in Japanese Laid-Open Patent Publication No. 63385/1983 and U.S. Pat. No. 4,657,866. However, so long as we carried out an animal cell culture test in a medium containing p-aminobenzoic acid, an animal cell growth enhancement effect could not be obtained.
Further, it is disclosed in Japanese Laid-Open Patent Publication No. 291796/1989 that, in production of human tissue plasminogen activator (tPA) by adherent cells, it is effective for increase of the productivity of single-strand tPA and prevention of detachment of the cells from the vessel wall or beads to add a p-aminomethyl benzoate in the medium.
On the other hand, it is disclosed in the WO93/22448 pamphlet subjected to International Publication on Nov. 11, 1993 that a culture broth where a useful protein was accumulated in a high concentration was obtained by culturing a human fetal nephric cell-derived 293 strain, in which a gene expressing the useful protein was introduced, in a medium comprising an e-RDF medium comprising a mixture of RPMI 1160 Medium, Ham's F-12 Medium and DULBECCO modified Eagle's Medium in 2:1:1 having added amino acids, glucose, etc. thereto, having added to the e-RDF medium insulin, transferrin, ethanolamine and sodium selenite as growth factors. However, when the example of the above WO93/22448 was checked, there could not be obtained any culture broth where the useful protein was accumulated in such a high concentration as described in the example, although the reason is not clear.
The main object of this invention is to provide low molecular compounds for addition to media for culture of animal cells, which can express, with reproducibility, an animal cell growth enhancement action.
Another object of this invention is to provide a method of proliferating animal cells or increasing the yield of a useful substance produced thereby, with reproducibility, by culturing the animal cells in a medium containing such a low molecular weight compound.
Other objects and characteristics of this invention will be apparent from the following description.


DISCLOSURE OF INVENTION

This invention provides a method of culturing animal cells in a medium characterized in that said medium contains a compound represented by the general formula: ##STR2## wherein, R denotes COOM or CHO, herein M denotes hydrogen, an alkali metal or a C.sub.1 to C.sub.3 alkyl group, and n is an integer of 1 to 3.
According to this invention is further provided a medium compositio

REFERENCES:
patent: 4657866 (1987-04-01), Kumar
Ohashi, et al. 1984. PNAS, vol. 81, pp. 7132-7136.
Pace, et al. 1967, Canadian Journal of Biochemistry, vol. 45 pp. 81-88.

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