EP0304358B1 - Röhrchen zur Aufbewahrung von kleinen Mengen, insbesondere von biologischen Substanzen, und dessen Verwendung bei einem Befruchtungsverfahren - Google Patents

Röhrchen zur Aufbewahrung von kleinen Mengen, insbesondere von biologischen Substanzen, und dessen Verwendung bei einem Befruchtungsverfahren Download PDF

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Publication number
EP0304358B1
EP0304358B1 EP88401903A EP88401903A EP0304358B1 EP 0304358 B1 EP0304358 B1 EP 0304358B1 EP 88401903 A EP88401903 A EP 88401903A EP 88401903 A EP88401903 A EP 88401903A EP 0304358 B1 EP0304358 B1 EP 0304358B1
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EP
European Patent Office
Prior art keywords
tube
plug
needle
region
sealing
Prior art date
Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
Expired - Lifetime
Application number
EP88401903A
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English (en)
French (fr)
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EP0304358A1 (de
Inventor
Robert Cassou
Joseph Henny
Maurice Cassou
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Individual
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Individual
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Publication date
Priority claimed from FR8710359A external-priority patent/FR2618452B1/fr
Priority claimed from FR8715741A external-priority patent/FR2623205B1/fr
Application filed by Individual filed Critical Individual
Publication of EP0304358A1 publication Critical patent/EP0304358A1/de
Application granted granted Critical
Publication of EP0304358B1 publication Critical patent/EP0304358B1/de
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Expired - Lifetime legal-status Critical Current

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    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61D—VETERINARY INSTRUMENTS, IMPLEMENTS, TOOLS, OR METHODS
    • A61D19/00—Instruments or methods for reproduction or fertilisation
    • A61D19/02—Instruments or methods for reproduction or fertilisation for artificial insemination
    • A61D19/022—Containers for animal semen, e.g. pouches or vials ; Methods or apparatus for treating or handling animal semen containers, e.g. filling or closing
    • A61D19/024—Tube-like containers, e.g. straws

Definitions

  • the present invention relates to a flake of the kind used for the storage of small quantities of substances, in particular biological substances, consisting of a segment of tube the two ends of which respectively define a first and a second end zone, the first end zone being equipped with a corresponding plugging means, said first.
  • such a flake is in the form of an elongated thin tube equipped, in one of said end zones, with a tripartite plug comprising two buffers of a fibrous material permeable to gases and which encloses a powder initially also permeable to gases and capable of transforming on contact with an aqueous liquid into an impermeable paste or gel by creating a sealed stopper which can nevertheless slide in the tube.
  • a tripartite plug comprising two buffers of a fibrous material permeable to gases and which encloses a powder initially also permeable to gases and capable of transforming on contact with an aqueous liquid into an impermeable paste or gel by creating a sealed stopper which can nevertheless slide in the tube.
  • the other end zone of the tube segment normally remains open for the introduction of the liquid to be preserved, generally by suction exerted through the tripartite plug.
  • Patent application DE-A-2 506 108 (POPOVIC) describes a flake plugged in one of its two end zones by another tripartite plug initially impermeable to gas but capable of forming a tight piston.
  • This tripartite plug has instead of pads made of a gas permeable fibrous material of the CASSOU cap, pads of a gas impermeable polystyrene foam, enclosing a powder similar to that of the CASSOU cap liquids are introduced into this flake by translation of the tripartite plug from one end to the other.
  • Said polystyrene foam pads have the serious drawback of considerably reducing their volume at the temperature of liquid nitrogen and of transforming into a layer of material which no longer closes the tube. Only the gel from the powder then ensures the separation of the interior of the straw with the exterior.
  • the present invention relates to the creation of a straw offering high protection against any risk of contamination while being suitable for simple, convenient and safe handling.
  • the present invention thus relates to a straw for the storage and handling of small quantities of biological substances, composed of a rectilinear segment of tube made of transparent polymer material comprising at its two ends first and second end zones equipped with plugging means respective watertight, characterized in that these sealing means each consist of a self-sealing compact elastomer plug, called ECA, capable of sealingly adhering to the internal wall of the tube and allowing a hollow needle to pass through it, the tightness being maintained both around the needle in place and in the plug after removal of that -this.
  • ECA self-sealing compact elastomer plug
  • the straw according to the invention when it is empty of any substance, already has at these ends first and second sealing means constituted by the two plugs of self-sealing elastomer.
  • ECA electrowetting-co-sealing plug
  • the withdrawal of the needle causes the filling, thanks to the elasticity of the elastomer, of the channel created by the needle in the stopper.
  • the two self-sealing plugs allow you to inject and remove any fluid from either end, by inserting a needle into each of the plugs.
  • Another object of the present invention relates to the use of a straw, where the first as the second sealing means is constituted by an ECA plug, in a process the in-vitro fertilization of at least one female gamete by a gamete male from a group of male gametes, the gametes being of human or animal origin.
  • This method consists in forming in the tube segment a plurality of contiguous liquid segments respectively adapted to constitute on either side of an intermediate connection zone, a zone of loading of male gametes near one of said terminal areas and a loading area of female gametes near the other terminal area.
  • This process has the advantage of being biomimetic, in that the sperm have to travel a certain distance in a small diameter tube before waiting for the oocytes to ensure fertilization.
  • the oocytes are reached first by the most mobile and vigorous sperm.
  • the straw of Figure 1 consists of a segment of cylindrical tube 1 open at its two ends 2, 3. These define respectively a first and a second end zone.
  • the first end zone contains, inside the tube 1, a tripartite plug 4.
  • This stopper 4 is constituted by a segment 5 of a special powder between two pads 6, 7 of a fibrous substance permeable to the fluid which is presented here in the form of a braided wick.
  • the special powder of segment 5, initially porous to gases, has the property of rapidly transforming on contact with an aqueous liquid into an impermeable paste or gel which adheres to the wall of the tube and hardens on contact with air, thus forming an airtight stopper. to fluids.
  • the prior art tube thus has only one stopper.
  • the filling of the segment 8 of the tube 1 with an aqueous liquid can only take place through the open end 3 because of the very function of the tripartite plug.
  • the straw of FIG. 2 also includes a tripartite plug 104 in a first end zone.
  • a segment 5 of special powder is inserted between two pads 106, 107 of polystyrene foam.
  • the quality of the foam gives this tripartite stopper 104 a function different from that of the tripartite stopper 4 in FIG. 1.
  • this stopper 104 is gas tight and essentially serves as a movable piston inside the straw.
  • FIG. 3 represents a straw according to the invention which, compared with the straw of the prior art, further comprises towards the edge of the first and second ends 2 and 3 of the tube 1, inside, plugs 9 made of compact elastomer .
  • This stopper has a length limited to a small fraction of the total length of the tube, for example one twentieth of it.
  • This plug 9 called ECA plug, consists of a compact elastomer which adheres to the inner wall of the tube and which has the property of being passed right through by a hollow injection needle by creating a sort of channel which grips the needle body tightly.
  • This momentary drilling allows communication the inside of the tube 1 with the outside via the plug 9 made of compact elastomer, and therefore of carrying out material transfers via the needle duct between the inside of the tube and the outside.
  • the compact elastomer will have a final Shore A hardness of between 20 and 50.
  • compact elastomer an elastomer without a cell, so as to avoid collapse and shortening of the plug at cryopreservation temperatures.
  • the compact elastomer can be a thermoplastic rubber chosen from the cariflexes TR (registered trademark of the Shell company) Solprene (registered trademark of the Philips company). Europrene Sol T (registered trademark of Anic) or a one- or two-component elastomer, vulcanizable at room temperature like Rhodorsil translucent silicone sealant (registered trademark of Rhône-Poulenc Corporation), Silopren (registered trademark of Bayer Company), Pourable Silicone (registered trademark of Rezolin Company) or Silbione RTV (Rhône-Poulenc) with two components.
  • Rhodorsil translucent silicone sealant registered trademark of Rhône-Poulenc Corporation
  • Silopren registered trademark of Bayer Company
  • Pourable Silicone registered trademark of Rezolin Company
  • Silbione RTV Silbione RTV
  • the elastomers are chosen to be non-cytotoxic and with no salting-out or absorption properties with respect to the biological medium introduced into the tube.
  • the plug 9 made of compact elastomer can be placed inside the tube by conventional plugging, using a preformed plug, by force-fitting inside the tube 1.
  • the tube is held during this operation in a support barrel which encloses it tightly to avoid any deformation or buckling.
  • Another preferred method consists in injecting inside the tube towards its end 3 a metered amount of a thermoplastic elastomer or vulcanizing cold.
  • FIG. 4 represents a straw where the first plugging means comprises a tripartite plug 4, and where a conventional hollow needle 18 has been installed, by perforation of the second plug ECA 9, at the factory.
  • a bicone "double male Luer” connects this conventional hollow needle to another conventional hollow needle.
  • the beveled tip of the needle 18 protrudes inside the tube the plug of a certain length. This overshoot is provided in the event of injection of liquid by the needle 18 to obtain the creation of a gas bubble whose length will be all the more important as the tip of the needle 18 advances inside. of the tube.
  • the aspiration by the end 2 of the tube creates a kind of piston of liquid which goes in magnification and which moves towards the tripartite stopper until reaching the powder of the segment 5, which stops the suction by clogging.
  • the gas is normally air, but may be another inert gas if the tube has been filled with another gas.
  • FIG. 5 represents a straw where the first plugging means comprises an ECA plug 19 as well as a tripartite plug 4 and the second plugging means an ECA plug 9 equipped with a needle 15 with male end, adapted to be connected to a female Luer cone of a conventional injection needle.
  • the tripartite plug 4 was moved during manufacture towards the inside of the tube to make room for the plug 19.
  • FIG. 6 represents an embodiment of the straw of FIG. 5 in which a conventional needle 22 which passes through the plug 19 and a needle 23 with male end in truncated cone without guard which have been installed in the factory crosses the plug 9.
  • FIG. 7 schematically represents an apparatus intended for the automatic positioning of the hollow needle with male end 15 or 23 in a straw according to the invention with guidance along the axis of the tube 1.
  • the apparatus consists of a part 24 which holds the tube 1 in a gutter and a part 25 which holds the needle 15 along the axis of the tube. A translation of 25 approaching 24 has pushed the needle 15 into the desired position.
  • a flake fitted as in FIG. 5 with an elastomeric plug 19 not equipped with a needle and an ECA plug 9 not equipped with a needle as in FIG. 3 allows the manufacture in the factory of a flake which contains between its two ECA caps 9, 19 an inert atmosphere which can be chosen in particular from air, carbon dioxide, nitrogen, rare gases, or even a culture medium confined between two gas bubbles.
  • the straw like that of FIG. 3 does not have a three-part stopper 4 or 104, it is possible to inject or remove any fluid through one or other of the ends 2, 3 of the tube 1 provided that the two ECA 9 stoppers are each pierced by a hollow needle.
  • the tearing of the needles from their ECA cap recreates a seal.
  • the needles are for single use and discarded after use under conditions defined according to the risks of contamination and toxicity.
  • the flakes according to the invention allow the storage in flakes of doses of any substance which is not chemically aggressive for the components of the tube, in initially liquid, gaseous, or particulate form.
  • the particles must be fine enough to be able to flow through the channel of a needle.
  • the flakes according to the invention withstand sudden transitions in both directions between -196 ° C and about 34 ° C. All the components of the flakes are also chosen to be able to withstand sterilization, in particular by irradiation.
  • the flakes are advantageously made, in a manner already known, in a crystal quality material with a wall thickness of approximately 10/100 mm, their transparency and optical quality is such that it is possible to look with a binocular magnifier. passes inside the chaff, especially in the case of gametes, to observe the early stages of embryo development.
  • One of the plugs namely A, is pierced by a bevelled hollow needle which serves as an exhaust orifice, the point of the needle barely emerging from the plug.
  • the other stopper namely B
  • the injection needle is removed from the stopper B.
  • the cap B is again pricked, this time passing the needle d about 15 mm, and a segment of the diluent medium is injected about 10 mm in length; this causes the outlet needle of the stopper A to leave a corresponding volume in the middle of Menezo.
  • a diluting medium for example F1 or Tris medium containing spermatozoa
  • the plug B is then re-drilled using a bevelled hollow needle whose tip barely exceeds the plug.
  • oocyte (s) With a needle and a small capacity syringe, a few drops of a diluent containing one or more oocytes are injected through the stopper A. Excess liquid escapes through the hollow needle of stopper B. This introduction of oocyte (s) into the tube can be checked by examination with a binocular microscope.
  • the tube thus filled is kept horizontal at a temperature in the region of 37 ° C, and the cleavage of the oocytes is checked by visual examination.
  • the embryos reach the stage of division 4 or 8, they are collected in a Petri dish by cutting the tube by a cutting tool perpendicular to the axis of the tube, that is to say in an area located outside the neighboring stopper embryos, either in an area opposite the stopper.
  • Each embryo collected can be directly implanted in a recipient woman, or frozen for conservation in a French straw in a medium containing a cryoprotector, such as glycerol, ethylene glycol or propylene glycol.
  • a cryoprotector such as glycerol, ethylene glycol or propylene glycol.
  • the embryo can be transferred into a straw, according to the present invention, using removable hollow needles.
  • the embryo is placed in its nourishing medium containing the cryoprotective and it is arranged, by successive injections, so that the embryo is found inside the tube in its medium in the form of a segment of liquid surrounded by an air bubble on each side.
  • Air bubbles act as shock absorbers during volume changes caused by liquid-solid and solid-liquid phase transitions during freezing and thawing. In addition, these air bubbles serve as a separator between different segments of different liquid which one does not wish to mix at the start.

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  • Health & Medical Sciences (AREA)
  • Life Sciences & Earth Sciences (AREA)
  • Veterinary Medicine (AREA)
  • Reproductive Health (AREA)
  • Engineering & Computer Science (AREA)
  • Wood Science & Technology (AREA)
  • Zoology (AREA)
  • Animal Behavior & Ethology (AREA)
  • General Health & Medical Sciences (AREA)
  • Public Health (AREA)
  • Apparatus Associated With Microorganisms And Enzymes (AREA)
  • Medical Preparation Storing Or Oral Administration Devices (AREA)

Claims (5)

1. Röhrchen zur Aufbewahrung und Handhabung von kleinen Mengen biologischer Substanzen bestehend aus einem geradlinigen Abschnitt (8) eines Röhrchens (1) aus durchsichtigem Polymerisatmaterial, das an seinen zwei Enden (2, 3) eine erste und zweite Endzone aufweist, die mit jeweiligen dichten Verschließvorrichtungen (9, 10) ausgerüstet sind, dadurch gekennzeichnet, daß diese Verschließvorrichtungen jede aus einem Stopfen aus einem kompakten selbstabdichtenden Elastomer vom Typ ECA bestehen, das dicht an der Innenwand des Röhrchens (1) haften kann und durch welches eine Hohlnadel hindurchgeschoben werden kann, wobei die Abdichtung sowohl um die eingeschobene Nadel wie nach deren Zurückziehung im Stopfen erhalten bleibt.
2. Röhrchen nach Anspruch 1, dadurch gekennzeichnet, daß das selbstabdichtende Elastomer ausgewählt ist aus den thermoplastischen Kautschuken und den bei Raumtemperatur vulkanisierbaren Silikonelastomeren.
3. Röhrchen nach einem der Ansprüche 1 und 2, dadurch gekennzeichnet, daß das selbstabdichtende Elastomer eine Shore-Härte zwischen 20 und 50 aufweist.
4. Verwendung eines Röhrchens nach einem der Ansprüche 1 bis 3 in einem Verfahren zur biomimetischen Befruchtung, wobei man im Abschnitt (8) des Röhrchens eine Mehrzahl von aneinanderstoßenden Flüssigkeitssegmenten bildet, welche auf der einen und anderen Seite einer dazwischenliegenden Verbindungszone eine Füllzone von männlichen Gameten in der Nähe der ersten Endzone und eine Füllzone von weiblichen Gameten in der Nähe der zweiten Endzone bilden können.
5. Verwendung nach Anspruch 4, dadurch gekennzeichnet, daß die Füllzone der männlichen Gameten vom ersten Stopfen durch ein Flüssigkeitssegment von gleicher Art wie die Verbindungszone getrennt ist, während die Füllzone der weiblichen Gameten an den zweiten Stopfen anstößt.
EP88401903A 1987-07-22 1988-07-22 Röhrchen zur Aufbewahrung von kleinen Mengen, insbesondere von biologischen Substanzen, und dessen Verwendung bei einem Befruchtungsverfahren Expired - Lifetime EP0304358B1 (de)

Applications Claiming Priority (4)

Application Number Priority Date Filing Date Title
FR8710359A FR2618452B1 (fr) 1987-07-22 1987-07-22 Paillettes pour la conservation de petites quantites de substances notamment biologiques
FR8710359 1987-07-22
FR8715741 1987-11-13
FR8715741A FR2623205B1 (fr) 1987-11-13 1987-11-13 Receptacle pour substances biologiques et procede de fecondation utilisant un tel receptacle

Publications (2)

Publication Number Publication Date
EP0304358A1 EP0304358A1 (de) 1989-02-22
EP0304358B1 true EP0304358B1 (de) 1992-03-04

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EP88401903A Expired - Lifetime EP0304358B1 (de) 1987-07-22 1988-07-22 Röhrchen zur Aufbewahrung von kleinen Mengen, insbesondere von biologischen Substanzen, und dessen Verwendung bei einem Befruchtungsverfahren

Country Status (2)

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EP (1) EP0304358B1 (de)
DE (1) DE3868782D1 (de)

Cited By (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US6213171B1 (en) 1999-01-19 2001-04-10 Imv Technologies Method and system for filling a straw with a liquid

Families Citing this family (8)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
FR2651793B1 (fr) * 1989-09-14 1991-12-06 Cassou Robert Tubes ou paillettes pour la conservation cryogenique d'echantillons biologiques tels que virus, et procede de remplissage.
FR2651676B1 (fr) * 1989-09-14 1991-11-29 Derien Jean Charles Paille d'aspiration pour prelevement de produits liquides mouillants, procede et moyens de realisation de cette paille.
FR2686247B1 (fr) * 1992-01-20 1999-05-07 Lecointe Pascal Paillette destinee a recevoir une substance biologique.
FR2753367B1 (fr) * 1996-09-19 1998-12-04 Lecointe Pascal Paillette de stockage d'une substance biologique liquide et son procede de remplissage
FR2762210B1 (fr) * 1997-04-21 1999-07-02 Instr Medecine Veterinaire Paillette avec un bouchon en fibre pleine microporeuse
FR2810535B1 (fr) * 2000-06-27 2002-10-11 Eurl Cryo Vet Procede de remplissage d'une paillette de stockage avec de la semence
FR2824255B1 (fr) * 2001-05-07 2004-02-06 Imv Technologies Paillette pour la conservation de petites quantites de substances, notamment de liquides biologiques
FR2886931B1 (fr) * 2005-06-14 2007-08-17 Cryo Bio System Sa Procede pour conditionner dans une paillette une dose predeterminee de substance liquide et dispositif pour le mettre en oeuvre

Citations (1)

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Publication number Priority date Publication date Assignee Title
FR2194453A1 (de) * 1972-08-04 1974-03-01 Alter Richard

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BE498210A (de) * 1949-09-20
LU38125A1 (de) * 1959-02-04
CH533542A (fr) * 1971-08-25 1973-02-15 Geneco Appareil pour remplir des tubes d'un liquide
DE2506108A1 (de) * 1975-02-13 1976-09-02 Peter Dipl Ing Popovic Einrichtung mit rohrfoermigem gebilde weitgehend konstanten querschnitts und mit darinnen beweglichem kolben
US4219912A (en) * 1978-10-10 1980-09-02 Baxter Travenol Laboratories, Inc. Injection site having thermoplastically sealed injection port
DE3347714A1 (de) * 1983-12-31 1985-07-11 Rehau Plastiks Ag + Co, 8673 Rehau Besamungshuelle fuer die kuenstliche besamung von tieren

Patent Citations (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
FR2194453A1 (de) * 1972-08-04 1974-03-01 Alter Richard

Cited By (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US6213171B1 (en) 1999-01-19 2001-04-10 Imv Technologies Method and system for filling a straw with a liquid

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EP0304358A1 (de) 1989-02-22
DE3868782D1 (de) 1992-04-09

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