Chemistry: molecular biology and microbiology – Spore forming or isolating process
Patent
1984-11-21
1988-11-22
Tarcza, John E.
Chemistry: molecular biology and microbiology
Spore forming or isolating process
4352403, 435948, C12N 500, C12R 191
Patent
active
047865997
DESCRIPTION:
BRIEF SUMMARY
The invention relates to animal cell culture media.
It relates more particularly to a synthetic medium for animal cell culture, particularly of epithelial cells or myeloma cells and their hybrids of hybridoma type, not containing serum and suitable both for primary culture of animal cells, and for production of cell lines from this primary culture or from another. It also relates to a primary culture process and a process for producing an animal cell line using such a medium.
For a certain number of years, intensive research has been carried out in order to perfect a culture medium enabling hepatic cell lines to be obtained having the characteristics of parenchymal hepatocytes, in order particularly to elucidate the regulatory mechanisms which control the proliferation and differentiation of hepatic cells.
Until now two types of hepatic cell cultures have been widely developed from cells obtained after dissociation of hepatic tissue by means of collagenase or trypsin.
It is known that the digestion of liver by collagenase provides non-proliferating parenchymal hepatocytes which can be kept several days, even several weeks, either in a culture medium supplemented with serum (Serum-Supplemented Medium: SSM), or in a culture medium without serum (Serum-Free Medium: SFM). These parenchymal cells manifest various functions of liver and consequently advantageously replace liver homogenates or tissue slices. However, the inevitable senescence of these hepatocytes does not allow long-term studies, which has prompted researchers to develop proliferating cultures of hepatic epithelial cells. Such cultures have been, in a first phase, obtained in media supplemented with serum (SSM), after digestion of hepatic tissue with trypsin or from primary cultures initiated after collagenase digestion. Although the growth of normal hepatic epithelial cells generally presents difficulties, particularly by reason of a too considerable development of fibroblastic cells, it has been possible to show that proliferating cultures of hepatic epithelial cells possess characteristics of hepatic parenchymal cells. However, the addition of serum to the culture medium introduces into the latter a very large number of components among which are particularly final products and effectors, that is to say regulators of the liver metabolisms which complicate, even render impossible, the use of cell lines obtained in serum-supplemented media, in particular in the case of certain studies such as those related to the biosynthesis of cholesterol, of neutral bile sterols and of bile acids.
To remedy, at least in part, the various drawbacks presented by cell lines obtained in serum-supplemented culture media, various authors have proposed replacing the serum by hormones and growth factors such as in particular insulin and "epidermal growth factor", whose presence until now was considered as indispensable for the development of cell lines.
Now, the dependence of numerous metabolic processes of liver with respect to hormones is well known. In consequence, the presence of hormones in the culture medium leads also to modifications or disturbances, even in certain cases to the impossibility of carrying out reliable studies on hepatic cell lines obtained in hormone-supplemented culture media.
In the case of the industrial use of established epithelial cell lines and of myeloma lines and their hybrids of the hybridoma type, the presence of serum or of unknown or poorly defined components leads to a great difficulty even to an impossibility, of purifying the one or more cell biology products for purposes of administration to man. In addition, the non-definition of the composition of the medium leads to prohibition of such human utilization of these biological products by the international and national health authorities (OMS, FDA, Ministry of Health, etc.).
The purpose of the invention is to provide an animal cell culture medium not presenting the drawbacks of known culture media.
Now it has been observed surprisingly and against all expectation, that it is po
REFERENCES:
patent: Re30985 (1982-06-01), Cartaya
patent: 4036693 (1977-07-01), Levine et al.
patent: 4440860 (1984-04-01), Klagsbrun
Kaighn, M. Edward, "Human Liver Cells", Tissue Culture Methods and Applications, Kruse, Jr., and Patterson, Jr. Ed., Academic Press, New York, 1973, pp. 54-58.
Maciag et al., "Hormonal Requirements of Baby Hamster Kidney Cells in Culture", Cell Biology International Reports, vol. 4(1), 1980, pp. 43-50.
Chessebeuf Martina L.
Padieu Prudent H.
Institut National de la Sante et de la Recherche Medicale
Tarcza John E.
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