Chemistry: molecular biology and microbiology – Animal cell – per se ; composition thereof; process of... – Culture medium – per se
Patent
1997-05-12
2000-09-05
Priebe, Scott D.
Chemistry: molecular biology and microbiology
Animal cell, per se ; composition thereof; process of...
Culture medium, per se
435406, 435408, 435384, 435385, 435386, 435387, 435373, 435349, 435391, 435392, C12N 500, C12N 506, C12N 516, C12N 502
Patent
active
061141682
DESCRIPTION:
BRIEF SUMMARY
The present invention relates to the obtaining of bird ES cells, especially to a method of culture and to a medium permitting the culture of these cells.
In effect, in the context of perfecting recombinant protein production technique, the development of a technique of transgenesis [sic] in domestic birds will have extremely important economic repercussions in two major applications: characters (resistance to certain diseases, growth performance features, and the like) albumin. possibility of producing proteins of interest in biological fluids or organisms (blood, milk, plants, etc.). The production of such proteins in domestic birds' eggs will certainly constitute a major technological advance in this approach, for several reasons: because their overabundance in these organisms has deleterious effects (for example erythropoietin, which induces pathological hyperglobulinemia in rabbits). Many of these proteins of interest do not display cross-activity with those of birds, thus permitting their overproduction in an avian organism without major pathological effect; mammals will come up against health problems associated with the presence in this [sic] species of latent organisms which are potentially pathogenic for man (lentiviruses, prions, etc.). This risk is very minimal, not to say almost nonexistent, in relation to pathogenic agents of domestic birds; number of proteins. For example, the major protein of birds'eggs, ovalbumin, represents 54% of the egg white proteins, equivalent to an average dry weight per egg of 2 grams of dry matter approximately. It is possible reasonably to conceive of producing per egg at least 10% of this mass as recombinant protein. The economic viability is seen to be very great if it is considered that a hen lays on average 2 eggs every three days, and this viability is seen to be much greater than that of large mammals if the much lower breeding costs of domestic birds are considered.
The production of transgenic birds is currently possible at an extremely high cost on account of its very low efficiency. In effect, in birds, the technique of microinjection of DNA into the egg is almost impossible. On the other hand, the use of the vector retrovirus system, the only efficient system to date, remains complex and will certainly come up against a reticence on the part of industrialists on health grounds.
A very great advance in the production of transgenic animals has been brought about in mice by the development of ES cell technology.
ES cells (embryonic stem cells) are totipotent embryonic cells capable of regenerating all the tissues of the embryo, including the germ tissue, after their injection into very early embryos. These cells may hence be considered to be Trojan horses for introducing new genetic information into an animal's genetic constitution. The possibility of culturing these cells in the long term in vitro affords the possibility of exercising numerous controls before their implantation in vivo. Moreover, these cells may be stored without limit in liquid nitrogen, which constitutes a potential for storage of a genetic constitution.
The use of ES cells nowadays constitutes the most promising approach in domestic birds for the efficient production of transgenic animals.
Recent work from a Canadian group (R. Etches at the Guelph station) has suggested that ES cells must exist in the bird embryo (Petitte et al., 1990). This group at [sic] succeeded in transplanting such cells into embryos and consequently producing animals whose genetic constitution is derived from that of the grafted cells. However, to date, it has not been possible for success to be achieved in culturing these cells in vitro; as a result, it has not been possible to use these cells to transfer a transgene in a stable manner. This is a major impediment to the exploitation of ES cell technology in birds. ES cells may be characterized by three essential types of criteria: (ECMA-7, SSEA-1 and SSEA-3, in particular).
To date, it has not been possible to obtain any culture of ES cells which are identified
REFERENCES:
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Pain Bertrand
Samarut Jacques
Centre National de la Recherche Scientifique "CNRS"
Ecole Normale Superieure de Lyon
Institute National de la Recherche Agronomique
Priebe Scott D.
Wilson Michael C.
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