Chemistry: molecular biology and microbiology – Spore forming or isolating process
Patent
1993-04-30
1996-07-02
Stone, Jacqueline M.
Chemistry: molecular biology and microbiology
Spore forming or isolating process
600 35, 600 33, 128738, 436 65, 436906, A61B 1743
Patent
active
055321550
DESCRIPTION:
BRIEF SUMMARY
BACKGROUND OF THE INVENTION
Since the birth in 1978 of Louise BROWN, the first child born as a result of in vitro fertilization, few major changes have taken place in the in vitro fertilization method.
Ovarian stimulation is now conventionally practiced on account of the ease with which oocyte retrieval can be programmed. The different ovarian stimulation procedures, clomiphene citrate - hMG, hMG alone, FSH - hMG and, more recently, the agonists of LH-RH associated with hMG, have considerably increased the number of oocytes obtained per retrieval (from 2.5 to 3.5 on average to 8 to 10 at the present time). This increase in the number of oocytes obtained has led to a larger number of embryos. In view of the risk of a multiple pregnancy caused by the transfer of more than three embryos, most assisted reproduction centers have acquired freezing apparatus to preserve the embryos and permit their secondary transfer replacement. Such stimulatory procedures are very expensive owing to the quantities of hormones used and the increased number of examinations (hormone doses and ultrasonic examinations) needed in order to monitor follicle growth.
The fertilization technique per se which is conducted in a laboratory using a CO.sub.2 incubator, a laminar flow hood, an inverted microscope, represents a major initial investment (see M. D. DAMEWOOD: In Vitro Fertilization:Insurance and financial considerations. Assisted Reproduction Review 1: 38, 1991). In addition the manipulation times, which are long due to the complexity of the technique, there is the need to monitor parameters such as CO.sub.2, temperature and relative humidity in the incubator. This correspondingly increases the cost of the procedure. This cost, which is high, estimated at $ 6,500 per attempt in the U.S.A. and approximately 15,000 Francs in France, makes it difficult to extend and disseminate this technique.
In addition the results which, at the best fertilization centers, are always below those for natural fertilization, namely 20% pregnancy per stimulated cycle the high cost, and the risks which result directly from this type of treatment (hyperstimulation, multiple pregnancies, storage and manipulation of frozen embryos) have prompted government agencies in the so-called developed countries to take steps to curtail the extension of assisted reproduction or procreation centers. Such steps run counter, of course, to the interests of sterile couples.
The applicant's experiments in in vitro fertilization, which were launched in 1979, have made it possible to develop a new procedure, which will be described hereinafter:
I. Intravaginal culture (IVC).
Reference is made, first of all, to international application WO 87/02879 filed on 7 Nov. 1986 and published on 21 May 1987, on which was based my U.S. Pat. No. 4,902,286 and to international application WO 88/08280 filed on 2 May 1988, on which was based my U.S. Pat. No. 5,084,004 and published on 3 Nov. 1988. All the features disclosed in these two my aforesaid U.S. Patents, shall be deemed to form part of the present description.
It was observed, in the first place, that the fertilization technique per se could be considerably simplified. Thus, I demonstrated, using the "CIVETE" technique, in French, "culture intra vaginale et transfert embryonnaire" or intravaginal culture and embryo transfer, that the addition of 5% CO.sub.2 enriched air was not necessary to maintain good culture conditions. However, to avoid any disturbance in the culture medium, it was necessary to fill the tube completely, without any air. Similarly, to avoid any major disturbance to the pH of the culture medium, the sperm concentration (responsible for metabolic consumption) was substantially reduced, 10,000 to 20,000 motile spermatozoa/ml, whereas the concentration is at least 50,000 mobile spermatozoa/ml in the conventional technique. This technique necessites fewer manipulations and requires no change in the culture medium.
In the conventional in-vitro fertilization technique, denuding or removal of the cumulus masa is carrie
REFERENCES:
patent: 5084004 (1992-01-01), Ranoux
Campell Bruce R.
Stone Jacqueline M.
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