Container for fertilization of human ovocytes in the absence of CO.sub.2 ...

United States Patent M

Ranoux

[li] Patent Number: 4,902,286 y [45] Date of Patent: Feb. 20, 1990

[54] CONTAINER FOR FERTILIZATION OF

HUMAN OVOCYTES IN THE ABSENCE OF
CO2-ENRICHED AIR

[76] Inventor: Claude Ranoux, 7 Rue des Grands Champs, 77330 Lesigny, France

[21] Appl. No.: 80,537

[22] PCT Filed: Not. 7,1986

[86] PCT No.: PCT/FR86/00378

§ 371 Date: Jul 6,1987

§ 102(e) Date: Jul. 6,1987 [87] PCT Pub. No.: WO87/02879

PCT Pub. Date: May 21,1987

[30] Foreign Application Priority Data

Nov. 8, 1985 [FR] France 85 16558

[51] IntCI." .. A61B 19/00

[52] U.S. CI 604/403; 600/33;

600/34; 435/296

[58] Field of Search 600/33, 34; 604/328,

604/330, 403; 128/341, 343, 769, 778; 435/1, 296, 299; 220/258

[56] References Cited

U.S. PATENT DOCUMENTS

1,877,766 9/1932 Kennedy 128/341

2,456,607 12/1948 Shaffer .

2,707,471 5/1955 Koff 128/341

2,818,064 12/1957 Leff 128/834

3,239,429 3/1966 Menolasino et al 435/296

3,805,784 4/1974 Alter .

3,875,012 4/1975 Dorn et al 435/296

4,300,544 11/1981 Rudel 128/832

4,380,997 4/1983 Leibo .

4,427,477 1/1984 Milgrom 128/837

4,533,345 8/1985 Louw .

4,555,037 11/1985 Rhees 220/258

4,579,823 4/1986 Ryder 435/296

4,598,045 7/1986 Masover et al 435/296

4,747,500 5/1988 Gach et al 220/258

4,761,379 8/1988 Williams et al 435/296

FOREIGN PATENT DOCUMENTS

43002 2/1930 United Kingdom 604/403

558998 1/1944 United Kingdom 604/403

2158093 11/1985 United Kingdom 435/296

Primary Examiner—C. Fred Rosenbaum
Assistant Examiner—Mark O. Polutta
Attorney, Agent, or Firm—Young & Thompson

[57] ABSTRACT

The invention relates to a container for fertilization of human ovocytes in the absence of CO2 which may be done in an incubuator at 37° C. or in the vaginal cavity. The container of cylindrical shape has smooth and rounded walls 1 whose thickness is 2 mm in the body 2 of the cylinder but only 1 mm at the neck 3. On the outer wall of the neck is seen a screwthread 4 permitting external screwing on of the plug 12 whose sides are smooth and rounded. The blind end 5 of the container is rounded, the other 6 is flat and has a round orifice 7, which orifice is closed by a fine membrane 9 permitting hermeticity of the tube in order to avoid disorders of the culture medium 8 present in the container as of manufacture. Above the liquid remains about 200 jal of gas 10. The inner walls 11 are smooth and rounded. After depositing the ovocytes and spermatozoa necessary for fertilization, and therefore after rupture of the membrane, hermeticity will be effected by two seals, after screwing on the plug, a circular one 13 at the neck of the tube, the other 14 on the endwall of the plug which will be flattened against the orifice of the tube 7 by a relief portion, located on the endwall of the plug 15, whose diameter is substantially that of the orifice. The container will be held in the posterior fornix of the vaginal cavity by a flexible ring comprised of a metal strip 17 sheathed in rubber 18 to which is fixed a rubber pouch 19 in which the container is placed. This device thus permits inter-vaginal, fertilization of human ovocytes in the absence of C02-enriched air, the vaginal cavity replacing the incubator conventionally used.

30 Claims, 2 Drawing Sheets

CONTAINER FOR FERTILIZATION OF HUMAN OVOCYTES IN THE ABSENCE OF CO2-ENRICHED AIR

5

This invention concerns a totally new procedure for fertilizing human ovocytes, making use of a device that we will describe.

In vitro fertilization of human ova is a very complex technique which permits a solution for cases of infertil- 10 ity of couples which up to then were irreversible. Since the first birth, of Louise Brown, in 1978, achieved by the Edwards team in England, thousands of children have been born worldwide through this technique.

The major concern of all teams working in vitro 15 fertilization has always been to obtain a simplification of the technique while maintaining or improving the results. Thus from the clinical standpoint there has appeared:

stimulation to obtain a plurality of ovocytes thereby 20 increasing the chances of success,

methods of ovarian puncture other than under coelioscopic control, namely under transvesical, then transvaginal and finally transurethral ultrasonic control which enabled general anesthesia to be eliminated and possible 25 CO2 toxicity to be avoided. From the biological standpoint the freezing of supernumerary embryos has permitted an improvement of results. Only the biological stage of fertilization per se has undergone but minimal changes. 30

It has remained very complex to date. Conventionally it involved aerobic or sterile culturing of embryos in a box or tube at 37° C. and under a 5% CO2 atmosphere. This requires non-hermetically sealed boxes or tubes with a risk of contamination by the surroundings. 35 Hence the need for a CO2 incubator (developed by "Testart") perfectly controlled at 37° C. and in CO2. This equipment is cumbersome and expensive.

Similarly, initially, after puncture, one proceeded to a l-to-4 hour maturation phase of the ovocytes in a cul- 40 ture medium usually enriched with human serum and, after this period, the fertilization of the ovocytes, changing the medium 18 to 24 hours after the stripping of the ova was effected. (This stripping involves the mechanical removal of the cumulus surrounding the 45 ovum to observe the stage of development.) The ovum was then transferred to a new medium which after 20 to 24 hours was again changed before transfer into the uterine cavity, which involved the use of more than 3 ml of culture medium per ovum and numerous manipu- 50 lations spread over 48 hours, which could be toxic to the ova.

The procedure that we have developed and whose scientific steps we will summarize is characterized by its simplicity and savings in time and money. 55

This procedure comprises fertilization of human ovocytes in the absence of C02-enriched air with the help of a fluidtight tube completely filled with culture medium that is placed in the vagina cavity which then serves as an incubator. Upon puncture of the ovocytes 60 which was delayed (so as to avoid the ovocyte maturation phase), they are placed in the container along with the spermatozoa necessary for fertilization which were previously prepared. Then the device with its holding means is placed in the vaginal cavity from where it will 65 be removed 44 to 48 hours later in order to reintroduce the ova in the uterine cavity by means of a "Frydman" catheter.

The invention will be better understood by consideration of the following description, taken in connection with the accompanying drawings, in which:

FIG. 1 is a cross sectional view of a container according to the present invention;

FIGS. 1A and IB are schematic views showing the methods of using the container of FIG. 1;

FIG. 2A is an elevational view of a pouch and ring for maintaining the container of FIG. 1 in place; and

FIG. 2B is an edgewise view, partly broken away, of the structure showing in FIG. 2A.

We are going to describe an embodiment of the container which constitutes the device (FIG. 1).

It is cylindrical tube of small dimensions, with smooth and rounded outer walls (1) in order to avoid any trauma to the vaginal mucosa. Its approximate dimensions are 4 cm long and 1.5 cm outer diameter; the thickness of the walls is 2 mm for the body (2) of the cylinder except in the region of the neck (3) where it is not more than only 1 mm, which gives an inner cavity of 1.1 cm in diameter and 3.8 cm long with an effective volume of about 3.2 cm3. This volume is suitable for the culture of 4 to 5 embryos. For a number of embryos greater than five, the volume of medium used may be increased to 5 cm3, the outer dimensions will then be 4.5 cm long and 1.7 cm outer diameter, the thickness of the wall remaining the same. Likewise shapes other than the cylindrical shape can be envisaged: round, archlike or pear-shaped depending on the configuration of the vagina.

On the outer wall of the body of the tube a marking surface must be provided for the patient's name. On the outer wall of the plug a screw limits aseptic defects.

Due to the decrease in the thickness of the wall of the neck, the outer edge of the plug, once it is screwed on, does not protrude beyond the wall of the tube. The blind end (S) of the tube is rounded. The upper end (6) of the tube adjacent the neck is flat and has at its center a round orifice (7) of about 4 mm in diameter. The ovocytes and the spermatozoa necessary for fertilization will be deposited through this orifice. The dimensions of the orifice has been purposely reduced in relation to the diameter of the tube in order to limit the communication between the culture medium and the surrounding atmosphere and therefore to minimize disorders caused thereby.

To reduce further septic risks, the medium disorders and the length of handling time, it seemed advisable to us to devise containers in which the culture (8) medium is placed in the tube at the time of its manufacture.

To avoid biochemical disorders to the medium between the time of manufacture and use, a hermetic closure of the tube will be effected by a thin membrane (9) covering the orifice. This membrane will be made so as to rupture easily when the ovocytes (22) and spermatozoa (24) are deposited in the tube by means of a glass "Pasteur pipette" (21, 23) or a throwaway plastic pipette end piece. Other types of sealed closure may be used such as a device with a valve.

The culture medium may be one of those conventionally used and marketed. The one that we use is the "I.N.R.A. de Menezo" medium marketed under the name "B2" by "A.P.I. Systeme". There will remain only a small volume (10) of gas of about 200 jxl between the surface of the medium and the membrane, corresponding to the volume of the ovocytes and spermatozoa transferred.