Classifications

• • A—HUMAN NECESSITIES
  • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
  • A61B—DIAGNOSIS; SURGERY; IDENTIFICATION
  • A61B17/00—Surgical instruments, devices or methods
  • A61B17/42—Gynaecological or obstetrical instruments or methods
  • A61B17/425—Gynaecological or obstetrical instruments or methods for reproduction or fertilisation
• • C—CHEMISTRY; METALLURGY
  • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
  • C12M—APPARATUS FOR ENZYMOLOGY OR MICROBIOLOGY; APPARATUS FOR CULTURING MICROORGANISMS FOR PRODUCING BIOMASS, FOR GROWING CELLS OR FOR OBTAINING FERMENTATION OR METABOLIC PRODUCTS, i.e. BIOREACTORS OR FERMENTERS
  • C12M23/00—Constructional details, e.g. recesses, hinges
  • C12M23/02—Form or structure of the vessel
  • C12M23/10—Petri dish
• • C—CHEMISTRY; METALLURGY
  • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
  • C12M—APPARATUS FOR ENZYMOLOGY OR MICROBIOLOGY; APPARATUS FOR CULTURING MICROORGANISMS FOR PRODUCING BIOMASS, FOR GROWING CELLS OR FOR OBTAINING FERMENTATION OR METABOLIC PRODUCTS, i.e. BIOREACTORS OR FERMENTERS
  • C12M23/00—Constructional details, e.g. recesses, hinges
  • C12M23/34—Internal compartments or partitions
• • C—CHEMISTRY; METALLURGY
  • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
  • C12M—APPARATUS FOR ENZYMOLOGY OR MICROBIOLOGY; APPARATUS FOR CULTURING MICROORGANISMS FOR PRODUCING BIOMASS, FOR GROWING CELLS OR FOR OBTAINING FERMENTATION OR METABOLIC PRODUCTS, i.e. BIOREACTORS OR FERMENTERS
  • C12M33/00—Means for introduction, transport, positioning, extraction, harvesting, peeling or sampling of biological material in or from the apparatus
  • C12M33/04—Means for introduction, transport, positioning, extraction, harvesting, peeling or sampling of biological material in or from the apparatus by injection or suction, e.g. using pipettes, syringes, needles
• • A—HUMAN NECESSITIES
  • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
  • A61B—DIAGNOSIS; SURGERY; IDENTIFICATION
  • A61B10/00—Instruments for taking body samples for diagnostic purposes; Other methods or instruments for diagnosis, e.g. for vaccination diagnosis, sex determination or ovulation-period determination; Throat striking implements
  • A61B10/0045—Devices for taking samples of body liquids
  • A61B10/0058—Devices for taking samples of body liquids for taking sperm samples

Definitions

• the present invention relates to a petri dish for selective collection of sperm used for selectively collecting highly motile sperm (motor sperm) in in vitro fertilization using the assisted reproduction technique (ART).
• ART assisted reproduction technique
• In vitro fertilization is performed on infertility cases caused by female factors such as obstruction of the fallopian tube, male factors such as a small number of good sperm, or both factors.
• the ovum and sperm collected from the plant are fertilized outside the body, cultured to some extent and then returned to the body.
• ART it is important to use morphologically normal motor spermatozoa extracted from semen for fertilization.
• Motor sperm removes seminal plasma by centrifugation using a density gradient body, concentrates semen, injects the semen into the lower part of the culture solution, and leaves it for a certain period of time. It is obtained by a method of collecting sperm that has been swim-up in the liquid (swim-up method). As shown in FIG. 10, this swim up method is applied to the pipette 94 as a moving sperm that swims up the sperm 93 present at the top of the culture medium 92 that is often performed in a general test tube 91. To collect. However, performing the swim-up method in a test tube in this manner has a poor recovery rate of motor spermatozoa, and it is not always possible to collect only motor sperm.
• a petri dish for the purpose of collecting motor sperm by the swim up method is known (for example, see Patent Document 1).
• This petri dish is for selectively collecting only the moving spermatozoa that have poured semen into a recess provided on the bottom of the petri dish, filled with the culture medium at the top, and swim up to the bottom of the petri dish from the recess.
• the motor sperm Although only the sperm present in the upper part of the culture medium after being left for a certain period of time is the motor sperm. Some say it is not. In other words, once the motility is high, the sperm force that once swims up to the upper part of the culture medium may move again to the lower part of the culture medium.
• a culture vessel provided with a plurality of culture compartments surrounded by a peripheral wall is known (see, for example, Patent Document 2).
• This container is used for culturing the culture by fixing the culture to the bottom of each storage compartment and filling the culture solution so as to cover all the storage compartments. If the container is used in a method of selecting a compartment into which semen is first injected and then collecting the sperm that has been transferred from the culture medium to another compartment as a motor sperm, compared to the petri dish described above. The efficiency of the work of collecting motor sperm is good.
• the culture container is not a container intended for selective collection of motor sperm
• the distance from one compartment to the other compartments varies, so the time required for sampling depends on which sperm moved to be a moving sperm.
• the amount of motile spermatozoa collected may be reduced.
• the width of the wall section that separates the central compartment from other compartments is not constant.
• the height of motility differs between motor sperm that moves with a partial force with a small width and motor sperm that moves with a partial force with a wide width.Therefore, the sperm with the mobility that can be collected depends on the width. It may remain in the compartment and the amount of motile sperm collected may be reduced.
• Patent Document 1 Japanese Unexamined Patent Publication No. 2003-289850
• Patent Document 2 Japanese Utility Model Publication No. 4 12898
• the present invention is a petri dish intended to collect motor sperm by the swim up method in normal in vitro fertilization, and can selectively collect only motor sperm in terms of sampling work and the amount of sampling.
• the purpose is to provide a petri dish.
• the present inventors have arranged a section into which semen is injected and a section to which sperm is moved to be adjacent to each other via a wall having a certain width at the upper end. So The present inventors have found that it is possible to provide a petri dish that can reliably and efficiently collect sperm having motility higher than that of the bell.
• a petri dish for selective collection of sperm having a bottom and a side wall, and having a plurality of receiving compartments therein, each containing compartment being lower than the side wall and having a constant upper end.
• the storage compartment is divided into one semen injection section into which semen is injected, and the semen injected into the semen injection section adjacent to the semen injection section through the wall.
• a petri dish for selective collection of sperm characterized by comprising one or more sperm transfer compartments in which sperm can move,
• the petri dish for selective collection of sperm is in the shape of the storage compartment itself, and the storage compartment is formed by dividing the interior of the petri dish for selective collection of sperm using the wall (1) Petri dish for selective collection of sperm according to any of (4),
• the storage compartment is provided in the center, and the storage compartment is arranged on the inside of the petri dish for selective collection of sperm on the wall and the bottom of the petri dish for selective collection of sperm.
• the petri dish for selective collection of sperm has a circular bottom, and the storage compartment is arranged around the semen injection compartment, which has a circular bottom surface arranged at the center of the bottom, and the semen injection compartment.
• a sperm transfer section having a bottom surface of the semen injection section and a concentric bottom surface
• the petri dish for selective collection of sperm of the present invention is arranged such that a section into which semen is injected (semen injection section) and a section to which sperm is moved (sperm transfer section) are placed adjacent to each other and semen is injected.
• sperm injection section a section into which semen is injected
• sperm transfer section a section to which sperm is moved
• the petri dish for selective collection of sperm according to the present invention has a sperm injection section and a sperm movement section adjacent to each other through a wall portion having a certain width at the upper end! Can be reliably collected.
• the level of sperm motility that can be collected can be appropriately set according to the height and width of the wall section that defines the storage compartment, or the angle of inclination provided on the upper end surface of the wall section.
• the petri dish for selective collection of sperm of the present invention can balance work efficiency and sperm collection efficiency depending on the number of sperm transfer sections, and has a suitable shape according to the state of semen injected into the semen injection section.
• a set petri dish can be used. That is, when the number of motile spermatozoa is small and semen is used, the number of sperm moving sections is large, and by using a sperm selective collection petri dish, precious motile spermatozoa can be collected as much as possible.
• the time required for collection can be shortened as much as possible by using a sperm selective collection petri dish with a small number of sperm transfer sections.
• FIG. 1 is a top view showing one embodiment of a sperm selective collection petri dish according to the present invention.
• FIG. 2 is a cross-sectional view of the sperm selective collection petri dish shown in FIG. 1 of the present invention.
• FIG. 3 is a cross-sectional view showing another embodiment of the petri dish for selective collection of sperm of the present invention.
• FIG. 4 is a cross-sectional view showing another embodiment of the sperm selective collection petri dish of the present invention.
• FIG. 5 is a top view showing still another example of the sperm selective collection petri dish of the present invention.
• FIG. 6 is a top view showing still another embodiment of the petri dish for selective collection of sperm of the present invention.
• FIG. 7 is a cross-sectional view of the sperm selective collection petri dish shown in FIG. 6 of the present invention.
• FIG. 8 is a top view showing still another example of the sperm selective collection petri dish of the present invention.
• FIG. 9 is a top view showing still another embodiment of the petri dish for selective collection of sperm of the present invention.
• FIG. 10 is an explanatory view showing a conventional method for selectively collecting motor spermatozoa.
• FIG. 1 is a top view showing an embodiment of the sperm selective collection petri dish of the present invention
• FIG. 2 is a cross-sectional view of the sperm selective collection petri dish shown in FIG. 3 and 4 are cross-sectional views showing other embodiments of the sperm selective collection petri dish of the present invention.
• 5, 8 and 9 are top views showing still other embodiments of the sperm selective collection petri dish of the present invention.
• FIG. 6 is a top view showing still another embodiment of the sperm selective collection petri dish of the present invention
• FIG. 7 is a cross-sectional view of the sperm selective collection petri dish shown in FIG.
• FIG. 10 is an explanatory diagram showing a conventional method for selectively collecting motor spermatozoa.
• the petri dish 1 for selective collection of sperm of the present invention comprises a bottom 2 and And side walls 3.
• a material constituting the petri dish 1 for selective collection of sperm a material that can be sterilized by electron beam and has high transparency, such as polystyrene, polyethylene, and polyamide, is preferably used.
• the petri dish for selective collection of sperm 1 is used in a state in which the culture compartment is filled so that the storage compartment described later is completely filled and the liquid level is above the upper surface of the storage compartment.
• the culture solution include a culture solution containing antibiotics such as cephalotin and Hepes as a buffer solution.
• the storage compartment is composed of one semen injection section 42 and one or more sperm transfer sections 43 adjacent to the semen injection section 42.
• the semen injection section 42 is a section for injecting concentrated semen after removing seminal plasma by centrifugation using a density gradient body in the swim up method of selectively collecting only motor sperm. .
• the semen injection section 42 is constituted by a bottom 2 of a sperm selective collection petri dish 1, a side wall 3 of the sperm selective collection petri dish 1, and a wall 41. Become.
• the volume of the semen injection section 42 is preferably 0.1 to 3. OmL.
• the sperm movement section 43 is a section used as a movement destination when the moving sperm in the semen injected into the semen injection section 42 moves in the culture solution.
• the sperm moving section 43 is constituted by a bottom 2 of a sperm selective collection petri dish 1, a side wall 3 of the sperm selective collection petri dish 1, and a wall 41.
• the sperm transfer section 43 is adjacent to the semen injection section 42 through the wall 41. Similar to the semen injection section 42, the sperm transfer section 43 also has a capacity of 0.1 to 3. OmL when the sperm selective collection dish 1 is used for the swim-up method. Is preferred.
• the height of the wall 41 that partitions the storage compartment is such that when the culture solution fills the inside of the petri dish 1 for selective collection of sperm, the liquid level is higher than the upper surface of the storage compartment , Set lower than side wall 3.
• the height of the wall 41 is such that the sperm in the semen injection section 42 is cultured. Since it becomes a barrier when moving in the liquid to the sperm moving section 43, it is determined as appropriate according to the mobility of the collected sperm. Considering the motility of the moving spermatozoa collected by the swim-up method in normal in vitro fertilization, the height of the wall 41 is preferably 1 to 5 mm.
• the height of the wall 41 is lower than lmm, there is a risk that even a sperm that is considered to have low mobility for use in in vitro fertilization may move to the sperm moving section 43, and the height of the wall 41 is 5 mm. If it is higher than that, the number of motile spermatozoa moving to the sperm moving section 43 becomes extremely small, which is not preferable.
• the upper end of the wall 41 needs to have a certain width in order to set the motility of the sperm moving from the semen injection section 42 to the sperm moving section 43 to a certain level or more.
• the width of the upper end of the wall 41 is also a barrier for movement of sperm, and is usually preferably 0.01 to 20 mm, more preferably 1 to 5 mm. If the upper end is constant, the width of the wall portion 41 may be set so that the lower end force also decreases toward the upper end. When the upper width of the wall portion 41 is set in the numerical range, it is preferable that the width of the lower end is set larger than that.
• the width of the upper end of the wall portion 41 is large, it becomes easier to visually recognize the sperm moving from the semen injection section 42 to the sperm moving section 43. However, if the width of the upper end of the wall 41 is larger than 20 mm, it takes time for the sperm to move from the semen injection section 42 to the sperm transfer section 43, and the sperm is placed on the wall 41 during the collection of the moving sperm It may remain. If the width of the wall 41 is smaller than 0.01 mm, the moving time of the sperm is short, but the moving sperm cannot be visually recognized. When the width of the wall portion 41 is large, the inside thereof may be a cavity as shown in FIG.
• the upper end surface of the wall portion 41 may be formed with a slope that increases from the semen injection section 42 side to the sperm movement section 43 side.
• the slope also serves as a barrier when sperm in the semen injection section 42 moves through the culture medium to the sperm transfer section 43, and can improve the fractionation efficiency of sperm in the wall 41. It is determined as appropriate according to the motility of the sperm to be collected.
• the fractionation efficiency of sperm refers to the probability of sperm that once moved from the semen injection section 42 side to the sperm transfer section 43 and further moved to the sperm injection section 42.
• the inclination of the upper end face of the wall 41 is The oblique is preferably 1 to 10 ° with respect to the horizontal direction. If the inclination of the upper end face of the wall 41 is smaller than 1 °, the fractionation efficiency of sperm is lowered.
• the wall portion 41 itself may be inclined with respect to the vertical direction of the bottom surface as shown in FIG.
• the inclination of the wall 41 is preferably an angular force of 10 to 80 ° formed by the wall 41 and the bottom surface, which is preferable from the viewpoint of improving the sperm fractionation efficiency.
• the shape of the storage compartment that is, the semen injection section 42 and the sperm transfer section 43 is not particularly limited as long as it does not hinder the injection of semen, the movement of motor sperm, and the collection of the motor sperm.
• a suitable shape of the accommodation compartment for example, as shown in FIGS.
• examples include a circular bottom surface and a polygonal bottom surface (not shown).
• each housing section may have a bottom surface and a wall portion arranged at right angles, or the wall portion may be arranged so as to be inclined outward from the vertical direction of the bottom surface. Furthermore, if the bottom surface and the wall portion are connected by a gentle curved surface and the edge of the bottom surface has a shape with no corners, it is easy to collect motor sperm.
• an identification symbol, a figure, or the like for distinguishing and distinguishing the semen injection section 42 and the sperm movement section 43 may be displayed in the storage section.
• the identification symbol or the like can be appropriately displayed on the bottom surface or wall 41 of each accommodation compartment, the bottom 2 or the side wall 3 of the sperm selective collection petri dish 1 and the like.
• a plurality of sperm moving sections 43 may be arranged in the sperm selective collection petri dish 1. Since the sperm transfer section 43 is disposed adjacent to the semen injection section 42 through the wall 41, the sperm injection section 42 is located at the center of all the storage sections.
• the petri dish 1 for selective collection of sperm shown in FIG. 5 has a sperm moving section 43 having a rectangular bottom surface on both sides of a semen injection section 42 having a rectangular bottom surface.
• the moving sperm injected into the semen injection section 42 can move into the sperm movement section 43 even if it moves to the left or right side. Therefore, the structure is more complex than that of the petri dish for selective collection of sperm 1 shown in FIGS. 1 and 2, but the sperm collection efficiency is good.
• a sperm moving section 43 having a bottom surface concentric with the bottom surface of the semen injection section 42 is disposed around the semen injection section 42 having a circular bottom surface. Being done.
• the moving sperm injected into the semen injecting compartment 42 can move into the sperm moving compartment 43 in any direction. Good.
• the sperm moving section 43 may be further divided into a plurality of storage sections by a wall 41 or an outer wall 44 described later, as shown in FIG. ,.
• the shape of the sperm selective collection petri dish 1 of the present invention is not particularly limited as long as the sperm selective collection petri dish 1 can easily grip the outer force of the side wall 3.
• the shape may be the shape of the containing compartment itself composed of the semen injection section 42 and the sperm transfer section 43 as shown in FIGS. 1 to 5, or as shown in FIGS.
• the bottom portion 2 in which the storage compartment is arranged at the center may be circular, or the bottom may be polygonal although not shown.
• the storage compartment is formed by dividing the interior of the sperm selective collection petri dish 1 using a wall 41.
• the storage compartment has an inner wall of the sperm selective collection petri dish 1 provided with a wall 41 and an outer wall 44 provided on the bottom 2. It is formed by using and partitioning. The height of the outer wall 44 is set higher than that of the wall portion 41 as shown in FIG.
• the sperm selective collection petri dish 1 shown in FIGS. 6 and 7 has a rectangular bottom section in the diameter direction of the bottom part 2 in the central part of the sperm selective collection petri dish 1 whose bottom part 2 is circular. It is provided so that it may extend.
• Each of the storage compartments, that is, the semen injection compartment 42 and the sperm transfer compartment 43 includes a bottom 2 of the sperm selective collection petri dish 1, a wall 41, and an outer wall 44 separately provided on the bottom 2.
• the petri dish 1 for selective collection of sperm shown in FIGS. 8 and 9 is also provided with a receiving section having a circular bottom at the center of the petri dish 1 for selective collection of sperm having a circular bottom 2.
• the liquid injection section 42 is composed of the bottom 2 and the wall 41 of the sperm selective collection petri dish 1, and the sperm transfer section 43 is separately provided with the bottom 2 of the sperm selective collection petri dish 1 and the wall 41. Consists of an outer wall 44.
• the sperm selective collection petri dish 1 in which the accommodation section is arranged in the central part has the same shape as the conventional petri dish and has good operability.
• the size is designed to be as small as necessary. Specifically, the outer diameter is about 30 to 60 mm. It is preferable.
• a convex portion may be further formed on the outer peripheral surface of the side wall 3.
• the convex portion has an effect as an anti-slip when the petri dish 1 for selective collection of sperm is also gripped by the outer force of the side wall 3.
• the convex portion may be provided in an annular shape over the entire outer peripheral surface of the side wall 3 or may be provided partially on the outer peripheral surface.
• the side wall 3 may be arranged in a vertical direction with respect to the bottom 2 or may be arranged so as to be inclined slightly outward with respect to the vertical direction of the bottom 2.
• the side wall 3 is inclined from the vertical direction of the bottom surface 2, the anti-slip effect when gripping the sperm selective collection petri dish 1 from the outside of the side wall 3 is improved, which is more preferable.
• the culture solution 45 is filled in the sperm selective collection petri dish 1.
• the amount of the culture solution 45 is preferably such that the liquid level is located slightly above the upper end of the wall 41.
• concentrated semen is injected below the semen injection section 42.
• the surface of the culture medium 45 may be covered with oil 46 such as paraffin to prevent the culture medium 45 from evaporating.
• the oil 46 may be injected either before or after injection of semen. Good.
• the sperm moves in the culture medium 45 through the semen injection section 42 from the semen injection section 42 to the sperm transfer section 43 through the wall 41 by its own mobility. If the wall 41 has a certain width, it is possible to visually recognize the movement of the sperm.
• sperm present in sperm transfer zone 43 is regarded as motile sperm and collected with culture medium 45 using a suction nozzle such as an injection needle or pipette.

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Cited By (11)

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• 2005
  • 2005-04-28 JP JP2005131351A patent/JP4586622B2/ja not_active Expired - Lifetime
• 2006
  • 2006-04-28 WO PCT/JP2006/309058 patent/WO2006118282A1/ja not_active Ceased

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