Chemistry: molecular biology and microbiology – Animal cell – per se ; composition thereof; process of...
Patent
1998-03-02
2000-10-10
Prats, Francisco
Chemistry: molecular biology and microbiology
Animal cell, per se ; composition thereof; process of...
435408, 435404, 435346, 435347, 435366, 435 11, 435 2, 600 33, 600 34, C12N 502, A61B 17425
Patent
active
061300861
DESCRIPTION:
BRIEF SUMMARY
TECHNICAL FIELD
The present invention relates to a culture medium composition for in vitro fertilization, in particular, applied to the culture of ova or early embryos which are fertilized ova or to the pretreatment of ova or sperm. In particular, the composition of the medium according to the present invention is effective for the stimulation of the growth and qualitative stabilization of early embryos and is suitable for the culture of early embryos.
BACKGROUND ART
The so-called in vitro fertilization and embryo transfer comprises fertilizing ova and sperm in vitro and then transplanting the developed embryos into a female body. Since the world's first case of a human birth was reported in England in 1978 by Edwards et.al., and along with recent progress in the developmental technology, this treatment has been rapidly and widely used in Japan, and it is now an indispensable treatment for sterility.
However, there are still few cases that lead to pregnancy by the method, and it can not be said that the method is completely established. This may be because of the lower fertility in sterile patients due to male factor, but the lower implantation rate of ova to be transplanted seems to be main cause ((Mori,Munehide et.al., Nippon sankahujin kagakukai zashi, v.45, p.397(1993); Cohen,J. et al., VIIIth World Congress on in vitro Fertilization and Alternate Assisted Reproduction Kyoto, Sep., 12-15(1993), World Collaborative Report(1991)).
In addition to the technical factors, a lowering in the quality of embryos during culture seems to be responsible for such lower implantation rates (Inoue, Masahito, Rinsho fujinka sanka, v.48, p.148(1994)). That is to say, ova of mammalia do not have substances that correspond to the albumin in the eggs of reptiles and birds, and therefore, the amounts of nutrient reserved in the mammalian ova are naturally low. Thus, in the early embryos of in vitro fertilization, nutrient factors must be taken up through the zona pellucida from the culture medium. However, chemically defined media such as Ham's F-10 medium, MEM (Minimum Essential Medium), Dulbecco's MEM and the like, which have been conventionally utilized in in vitro fertilization, were not originally composed for the purpose of in vitro fertilization, but they are the media used conventionally in tissue culture or their modified media, therefore it cannot be said that they are the optimal media for the early embryos regarding the nutrient composition.
Recently, HTF medium (Human Tubal Fluid Medium) has been developed as nutriologically suitable medium for human in vitro fertilization, with a composition approximating to the electrolyte of human oviduct fluid (Quinn,P. J. et al., Fertility and Sterility, v.44, p.493(1982)). The medium is commercially available and replaces Ham's F-10 medium that was used predominantly so far. However, because the HTF medium only contains electrolytes as the main components and glucose as an energy source, the HTF medium shows no improvement over the Ham's F-10 medium containing amino acids, as regards the nutrient composition. In fact, despite the use of this medium, the implantation rate can not be enhanced though any substantial improvement in the problem of the lowering in quality of embryos.
In order to make up for this disadvantage, a method has been utilized in which the embryos are fed by adding the female serum which has been inactivated by heat treatment to the medium. The serum contains growth factors and the like, in addition to proteins, carbohydrates, lipids, vitamins and minerals as five nutrients which are essential factors in animal cell culture. For reason, the serum is added during embryo culture.
However, it has been reported that such serum is not always needed in the in vitro fertilization-embryo transfer process (Menezo,Y. et al., Fertility and Sterility v.42, p.750(1984)) and that on the contrary the growth of embryos may be suppressed by the addition of serum (Mehita, et al.Biology of Reproduction v.43, p.600 (1990)). Also, the serum itself is troublesome to coll
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Araki Hiromasa
Kishi Yuichiro
Nakazawa Teruki
Ohashi Kazutomo
Shinoda Sanji
Fuso Pharmaceutical Industries Ltd.
Prats Francisco
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