 |
US
Patent & Trademark Office |
| Patent
Number |
5,908,380 |
| Title |
Compartmentalized
Zavos sperm swim-up column |
| Inventor(s) |
Panayiotis
M. Zavos;
Panayota N. Zarmakoupis-Zavos |
| Date
Granted |
June
1, 1999 |
| United States Patent |
5,908,380 |
| Zavos ,
et al. |
June 1, 1999 |
Compartmentalized Zavos sperm swim-up column
Abstract
The invention relates to a sterile, disposable, compartmentalized
Zavos swim-up column for the swimming-up of spermatozoa, at
desired levels of dilution in a media, and a swimming down
of spermatozoa into compartments within the column, as well
as to a method for the harvesting of those semen samples having
particular sperm morphology, motility, progressive motility,
speed, sperm concentration, fertilization potential, and a
sex ratio. The device comprises a hollow, vertically supported
column, having a closed lower end, and an open upper end.
A lowermost or first conical member is arranged at the lowermost
end of the column, the lowermost conical member having a lowermost
periphery in sealing engagement with the bottom of the column.
The first conical member has inclined side walls and a truncated
uppermost portion defining a peripheral edge which is open
within into the column. A second conical member having a lowermost
periphery is also attached to the side walls of the column
in a sealing manner therebetween. The second conical member
also has tapered walls and a truncated uppermost portion defining
a peripheral edge which is open to the inside of the column.
The area between the inclined walls of each of the conical
members and the inner wall of the column define a periconical
area which comprises a compartment for collecting sperm prior
to harvesting thereof.
| Inventors: |
Zavos; Panayiotis M.
(2413 Vince Rd., Nicholasville, KY 40356); Zavmaboupes-Zavos;
Panayota N. (2413 Vince Rd., Nicholasville, KY 40356) |
| Appl. No.: |
005802 |
| Filed: |
January 12, 1998 |
| Current U.S.
Class: |
600/33 |
| Intern'l Class:
|
A61D 007/00 |
| Field of Search:
|
600/33-35 128/897-98
604/403,405,406,410,906 435/2 |
References Cited [Referenced
By]
U.S. Patent Documents
Primary Examiner: Lacyk; John P.
Attorney,
Agent or Firm: Halgren; Don
Claims
We claim:
1. A sterile, disposable, compartmentalized Zavos swim-up column
for the swimming-up of spermatozoa, at desired levels of dilution
in a media, for the production of semen samples having particular
sperm morphology, motility, progressive motility, speed, sperm
concentration, fertilization potential, and a sex ratio, comprising:
a hollow, vertically supported column, having a closed lower
end, and an open upper end; and
a lowermost or first conical member arranged at the lowermost
end of said column, said lowermost conical member having a lowermost
periphery in sealing engagement with the lowermost end of said
column, said first conical member having inclined side walls
and a truncated uppermost portion defining a peripheral edge
and opened within said column; said area between said inclined
walls of said conical members and said inner walls of said column
defining a periconical area which comprises a compartment for
collecting sperm prior to harvesting thereof.
2. A sterile, disposable, compartmentalized Zavos swim-up column,
as recited in claim 1, including
a second conical member having a lowermost periphery attached
to side walls of said column in a sealing manner therebetween,
said second conical member having tapered walls and a truncated
uppermost portion defining a peripheral edge and open to the
inside of said column, said area between said inclined walls
of said conical members, and said inner walls of said column
defining a periconical area which comprises a compartment for
collecting sperm prior to harvesting thereof.
3. A sterile, disposable, compartmentalized Zavos swim-up column,
as recited in claim 1, including:
a sealable cap arranged for closing said open end of said column.
4. A sterile, disposable, compartmentalized Zavos swim-up column,
as recited in claim 2, including:
a further conical member arranged within said column, said further
conical member defining a further periconical compartment or
area between said inclined walls of said conical member and
the walls of said column, for the collection of sperm therein,
until harvesting thereof.
5. A method for the collection and harvesting of sperm samples
of several varying characteristics, comprising the steps of:
providing a column having a closed lower end and an open upper
end;
placing a first truncated conically shaped member at said lower
end of said column;
depositing a sample of sperm within said column within said
first conically shaped member;
filling said column with an isolation media above the upper
edge of said truncated conically shaped member;
permitting said sperm to swim up through said media about said
conical member and to swim down into a periconical compartment
between of said conical members and the walls of said column;
and
inserting a harvesting device to withdraw media and certain
sperm from a periconical compartment outwardly of said conical
member.
6. The method for the collection and harvesting of sperm samples
of several varying characteristics as recited in claim 5, including
the step of:
placing a second truncated conically shaped member in said column,
above said first conically shaped member.
7. The method for the collection and harvesting of sperm samples
of several varying characteristics as recited in claim 5, including
the step of:
inserting a harvesting device to withdraw media and certain
sperm from a further periconical compartment outwardly of a
further one of said conical members.
8. The method for the collection and harvesting of sperm samples
of several varying characteristics as recited in claim 6, comprising
the steps of;
arranging a third truncated conically shaped member within said
column, above said second conically shaped member; and
filling said isolation media above the uppermost edge of said
third conically shaped member, to provide sperm within said
first conically shaped member yet a further compartment in which
to swim up and swim down into.
Description
BACKGROUND OF THE INVENTION
1. Field of the Invention
This invention relates to an apparatus and a method of processing
semen and allowing its associated spermatozoa to become segregated
to permit subsequent selection thereof, to increase the number
of motile and morphological samples to be recovered therefrom.
2. Prior Art
A number of methods have been proposed for the processing of
semen so as to increase its quality and quantity of spermatozoa
population of a given semen sample. With improved quality and
quantity of such spermatozoa, the probability of success of
Artificial Insemination (AI), In Vitro Fertilization (IVF),
Intracytoplasmic Sperm Injection (ICSI), and other related Assisted
Reproductive Clinical Technologies (ARTs), may be improved.
Some of the methodologies include washing and storage of the
spermatozoa in media, and the "swim-up" procedure, a technique
that takes advantage of the swimming abilities of a small percentage
of spermatozoa within a given sperm population.
One attempt at the improvement of the collection of spermatozoa
is shown in U.S. Pat. No. 3,894,529 to Shrimpton, entitled,
"Method and Means For Controlling the Sex of Mammalian Offspring
And Product Therefor". This patent discloses a method of separating
X and Y sperm so as to control the sex of a desired offspring
by inseminating an egg with an X or a Y sperm. The sperm is
separated by applying a buoyant force to a mixture of sperm
in the nutrient media so that the separation occurs according
to the density of the sperm.
U.S. Pat. No. 4,759,344 To Wang, entitled, "Wang's Tubules For
Sperm Preparations Used For IVF-ET-GIFT and Artificial Inseminations",
discloses a method and an apparatus for separating sperm having
the greatest motility. The sperm are allowed to swim upward
in a column, where they may be collected to provide the strongest
samples for insemination.
Processes such as these are relatively simple, however they
are often inefficient and maybe susceptible to technician error
and/or to laboratory variations.
Further, none of these techniques allow the use of the "swim-up/swim-down"
principle nor do they permit selectivity based on the various
characteristics such progressive motility, morphological characteristics,
X or Y bearing sperm ratio and the like.
It is an object of the present invention, to provide a method
in apparatus for the selection of a desired spermatozoa population
from a given semen sample, to maximize the probability of success
when that semen sample is utilized for artificial insemination,
in vitro fertilization, intracytoplasmic sperm injection, and
other related assisted reproductive clinical technologies.
It is yet a further object of the present invention, to provide
a method and an apparatus that will minimize the likelihood
of technical error and laboratory variations, and permit the
selection of a particular population of spermatozoa.
It is yet still a further object of the present invention, to
provide a method in apparatus to take advantage of the "swim-up/swim-down"
principle and to permit the entrapment of various classifications
of spermatozoa based on their various characteristics such as
progressive motility, morphological characteristics, X or Y
bearing sperm ratio and the like.
BRIEF SUMMARY OF THE INVENTION
The present invention relates to an apparatus and a method for
processing semen and allowing its spermatozoa to be selectively
sampled, permitting an increase in the number of motile and
morphologically normal spermatozoa in the recovered fractions.
The apparatus comprises a generally vertically disposed, hollow
column having a planar or flat lowermost end. The column defines
a fluid-tight closure having an upper open end. A closure cap
is removably securable to the open upper end of the column.
A first or a lowermost truncated conical member having a wide
lowermost end and a narrower uppermost end is arranged at the
bottom of the column. The first truncated conical member has
a lowermost periphery which sealingly engages the lowermost
side wall portion of the column, in a sealing relationship therewith.
The walls of the first truncated conical member are arranged
at an angle up between about 45 to about 60 degrees with respect
to the longitude lower axis of the column. The area radially
outwardly of the longitudinal axis and the tapered walls of
the conical member is defined as the periconical area. The upper
end of the truncated conical member is opened to the chamber
defined by the interior walls of the column.
A second truncated conical member in a further embodiment, is
arranged longitudinally above the lowermost conical member.
The second conical member has a lowermost periphery that engages
and is in a sealed relationship with the walls of the column.
The second truncated conical member has an upper periphery through
which it is open (it is open at both ends), to the chamber defined
by the inside walls of the column. The area between the walls
of the column and the outer side of the second conical member
is defined as the periconical area or compartment number two.
The lowermost periphery of the second truncated conical member
is preferably disposed above the peripheral opening at the truncated
portion of the first conical member.
A third and uppermost truncated conical member may be arranged
in a mating relationship with the inside of the walls of the
column, the widest portion of the conical member being in a
sealed engagement with those inside walls of the column at a
location spaced above the upper end of the second conical member.
The third truncated conical member has an uppermost opening,
which is opened to the inside of the chamber defined by the
walls of the column. The space between the inside walls of the
column and the tapered walls of the third truncated conical
member is defined as the periconical area or compartment number
three.
The area above each respective uppermost periphery or truncated
portion of each conical member is defined as the epiconical
area of its respective adjacent conical member. The first conical
member is open only at its uppermost end. The second conical
member and the third conical member are each open at both their
lower and uppermost ends. Additional truncated conical members
may be added in a manner similar to the second and third conical
members, to increase the ability of the apparatus to discriminate
farther and to improve its accuracy when it is deemed necessary.
The method of utilization of the present invention involves
the placement of a semen specimen, (after it has been evaluated
for volume, sperm count per milliliter, motility, and presence
of debris), within the chamber defined by the lowermost conical
member through its opened upper end. The upper surface of the
semen sample should be brought to the same level as the upper
periphery of the open end of the lowermost or first conical
cylinder. Thereafter, a known volume of isolation media (i.e.
"Ham's F-10") is drawn to fill the periconical and the epiconical
areas of the first conical member, if only one such member is
utilized within the column, or into the epiconical area of the
second and third conical members, or more, depending on the
number of conical members used within the inventive device.
The isolation media must at least be filled above the lip of
the upper end of the particular conical member over which the
sperm is expected to swim to permit the "swim-up/swim-down"
process to occur. The column then has a closure cap that may
be then gently placed thereon, and the column and its contents
are allowed to sit in a controlled environment for about a period
of time, (for example, about one hour).
During that period of time, healthy, motile spermatozoa gradually
rise or swim up from the holding chamber within the lowermost
or first conical member, through the media, and into the epiconical
area of that first conical member. Certain spermatozoa will
subsequently swim down into the periconical area (the first
compartment) into the media surrounding the lowermost or first
conical member.
Certain spermatozoa specimens will gradually rise and swim-up
through the upper end of the second conical member and other
spermatozoa specimens will swim-up through the upper end of
the third (or higher) conical member. Certain of such specimens
will settle in particular periconical areas between the walls
of the middle or second conical member and the column, and the
walls of the third (or higher) conical member and the inside
wall of the column, (if more than one such conical member is
used), each respective "compartment" or periconical area each
defining a reservoir for a particular characteristic spermatozoa
therewithin.
At the end of this gradual-rise period, the over-layer of the
media in and around the top of the semen and within the respective
conical members for each individual compartment, is removed.
The particular spermatozoa entrapped in the media within that/those
periconical areas (compartments) are used for further cases
of assisted reproductive technologies. Such harvesting or removal
of the spermatozoa/media from the compartments within the column
at the end of the incubation period is done by placing a harvesting
instrument, such as a needle with a syringe, or a pipette, into
the respective periconical areas of each conical member, and
gently aspiring the media until almost all of the overlayed
media is removed each time. The gradual-rise specimen for each
compartment may be used at the desired level of dilution in
a media of choice, in a centrifuge operation. The level of sperm
dilution, or the sperm concentration in the resuspended preparation,
shall be determined by the clinical reproductive purpose for
that particular spermatozoa. Such gradually-risen spermatozoa
may be thus utilized for In Vitro Fertilization (IVF), Artificial
Insemination (AI), Intrauterine Insemination (IUI), Gamete Intra-Fallopian
Transfer (GIFT), Zygote Intra-Fallopian Transfers (ZIFT), Intra-Cytoplasmic
Sperm Injection (ICSI), sex selection (shifting the sex ratio
via the use of an X or Y bearing sperm fraction) and other similar
or related clinical assisted reproductive procedures or technologies.
Thus the invention comprises a sterile, disposable, compartmentalized
Zavos swim-up column for the swimming-up/swimming-down of spermatozoa,
at desired levels of dilution in a media, for the production
of semen samples having particular sperm morphology, motility,
sperm concentration, fertilization potential, and a sex ratio,
comprising a hollow, vertically supported column, having a closed
lower end, and an open upper end. A lowermost or first conical
member is arranged at the lowermost end of the column. The lowermost
conical member has a lowermost periphery in sealing engagement
with the lowermost end of the column. The first conical member
has inclined side walls and a truncated uppermost portion defining
a peripheral edge opening within the column.
A second conical member having a lowermost periphery is attached
to side walls of said column in a sealing manner therebetween.
The second conical member has tampered walls and a truncated
uppermost portion defining a peripheral edge. The second conical
member is open to the inside of said column. The area between
the inclined walls of the conical members, and the inner walls
of the column define a periconical area that comprises a compartment
for collecting sperm prior to harvesting thereof. A sealable
cap is arranged to close the open end of the column. A further
conical member is arranged within the column, the further conical
member defining a further periconical compartment or area between
the inclined walls of the conical member and the walls of the
column, for the collection of sperm therein, until harvesting
thereof.
The invention includes a method for the collection and harvesting
of sperm samples of several varying characteristics, may comprise
the steps of providing a column having a closed lower end and
an open upper end, placing a first truncated conically shaped
member at the lower end of the column, placing a second truncated
conically shaped member in the column, above the first conically
shaped member, depositing a sample of sperm within the column,
within the first conically shaped member, filling the column
with an isolation media to a level above the upper edge of the
second truncated conically shaped member, permitting the sperm
to swim up through the media about the conical member and to
swim down into a periconical compartment between one of the
conical members and the walls of the column, and inserting a
harvesting device to withdraw media and certain sperm from a
periconical compartment outwardly of one of the conical members.
The method may include the steps of inserting a harvesting device
to withdraw media and certain sperm from a further periconical
compartment outwardly of a further one of the conical members,
arranging a third truncated conically shaped member within the
column, longitudinally above the second conically shaped member;
and filling the isolation media to a level above the uppermost
edge of the third conically shaped member, to provide sperm
within the first conically shaped member yet a further compartment
in which to swim up and swim down into. Additional conically
shaped members may be placed in the column, on top of and spaced
from the third truncated conical member in a similar configuration
to the second and third conical members, to increase the ability
of the apparatus to further discriminate and improve its accuracy
in selecting higher quality sperm whenever deemed necessary.
BRIEF DESCRIPTION OF THE DRAWINGS
The objects and advantages of the present invention will become
more apparent when viewed in conjunction with the following
drawings, in which:
FIG. 1 is a side elevational view, in section, of a Zavos type
swim-up column constructed according to the principles of the
present invention;
FIG. 2 is a view similar to FIG. 1, showing placement of a sample
of sperm in the bottom of a cylinder;
FIG. 3 is a view similar to FIG. 2, showing the gradual-rise
of the sperm over a period of time;
FIG. 4 is a view similar to FIG. 3, showing a harvesting of
sperm from a top compartment;
FIG. 5 is a view similar to FIG. 4, showing a harvesting of
sperm from an intermediate compartment; and
FIG. 6 is a view similar to FIG. 5, showing a harvesting of
sperm from a lower
DESCRIPTION OF THE PREFERRED EMBODIMENTS
Referring now to the drawings in detail, and particularly to
FIG. 1, there is shown the present invention relates to an apparatus
and a method for processing semen and allowing its spermatozoa
to be selectively sampled, permitting an increase in the number
of motile, progressively motile and morphologically normal spermatozoa
in the recovered fractions.
The apparatus comprises a generally vertically disposed, hollow
column 10 having a planar or flat lowermost end 12. The column
10 defines a fluid-tight closure having an upper open end 14.
A closure cap 16 is removably securable to the open upper end
14 of the column 10.
A first or a lowermost truncated conical member 20 having a
wide lowermost end 22 and a narrower uppermost end 24 is arranged
at the bottom 12 of the column 10. The first truncated conical
member 22 has a lowermost periphery 26 which snuggly engages
the lowermost inner sidewall portion of the column 10, in a
sealing relationship therewith. The walls 28 of the first truncated
conical member 20 are arranged at an angle "A" of between about
45 to about 60 degrees with respect to the longitudinal axis
"L" of the column 10. The area radially outwardly of the longitudinal
axis and outwardly of the tapered walls 28 of the first conical
member 20 and within the walls of the column 10 is defined as
the periconical area "P1". The upper end 24 of the first truncated
conical member 20 is opened to the chamber defined by the interior
walls of the column 10, the area thereabove being defined as
its epiconical area "E1".
A second truncated conical member 30 in a further embodiment,
may be arranged longitudinally above the lowermost conical member
20, as shown in FIG. 1. The second conical member 30 has a wide
lowermost peripheral portion 32 that engages and is in a sealed
relationship with the walls of the column 10. The second truncated
conical member 30 has an upper peripheral portion 34 through
which it is open (open at both ends), to the chamber defined
by the inside walls of the column 10. The area between the walls
of the column 10 and the outer side of the second conical member
30 is defined as the periconical area "P2", as compartment number
two. The lowermost periphery 32 of the second truncated conical
member 30 is preferably disposed above the peripheral opening
24 at the truncated portion of the first conical member 20,
designated "E1".
A third and uppermost truncated conical member 40 is arranged
in a mating relationship with the inside of the walls of the
column 10, as shown on FIG. 1, the lowest and widest periphery
42 of the third conical member 40 is in a sealed engagement
with those inside walls of the column 10 at a location spaced
above the upper end 34 of the second conical member 30. The
third truncated conical member 30 has an uppermost opening 44,
which is opened to the inside of the chamber defined by the
walls of the column 10. The space between the inside walls of
the column 10 and the tapered walls 46 of the third truncated
conical member 40 is defined as the periconical area "P3" for
compartment number three.
The area above each respective uppermost periphery or truncated
portion of each conical member is defined as the epiconical
area E1, E2 and E3, for its respective adjacent conical member
20, 30 and 40, respectively. The first conical member 20 is
open only at its uppermost end 24. The second conical member
30 and the third conical member 40, and any additional such
members, are each open at both their lower and uppermost ends.
The method of utilization of the present invention involves
the placement of a semen specimen, (after it has been evaluated
for volume, sperm count per milliliter, motility, and presence
of debris), within the chamber defined by the lowermost conical
member 20 through its opened upper end 24. The upper surface
of the semen sample should be brought to the same level as the
upper periphery of the open end 24 of the lowermost or first
conical member 20, as may be seen in FIG. 2. Thereafter, a known
volume of isolation media (i.e. "Ham's F-10") is drawn to fill
the periconical and the epiconical areas of the second and third
conical members 30 and 40, and slightly above, as may be seen
in FIG. 3. The column 10 then has a closure cap 16, which is
then gently placed thereon, and the column 10 and its contents
are allowed to sit in a controlled environment for a set period
of time, (for example, about one hour). In any case, the time
set will be proportional to the number and quality of sperm
in the semen to be processed.
During that period of time, healthy, motile spermatozoa gradually
rise or swim up from the holding chamber within the lowermost
or first conical member 20, through the media, and into the
epiconical area "E1", of that first conical member. Certain
spermatozoa will subsequently swim down into the periconical
area (the first compartment P1) into the media surrounding the
lower-most or first conically shaped member.
Certain spermatozoa specimens will gradually rise and swim-up
through the upper end of the second conical member and other
spermatozoa specimens will swim-up through the upper end of
the third conical member. Certain of such specimens will settle
in particular periconical areas between the walls of the middle
or second conical member and the column, and the walls of the
third cone member and the inside wall of the column, each respective
"compartment" or area each defining a reservoir for a particular
class of spermatozoa therewithin.
At the end of this gradual-rise period, the over-layer of the
media in and around the top of the semen and within the respective
conical members for each individual compartment, is removed.
The particular spermatozoa entrapped in the media within those
periconical areas (compartments) are used for further cases
of assisted reproductive technologies. Such harvesting or removal
of the spermatozoa/media from the column at the end of the incubation
period is done by placing a harvesting instrument, such as a
needle with a syringe, or a pipette into the respective periconical
areas (P3, P2 and P1) of each compartment, and gently aspiring
the media until almost all of the overlayed media is removed
each time. The gradual-rise specimen for each compartment may
be used at the desired level of dilution in a media of choice,
in a centrifuge operation. The level of sperm dilution, or the
sperm concentration in the resuspended preparation, shall be
determined by the clinical reproductive purpose for that particular
group of spermatozoa. Such gradually-risen spermatozoa may be
thus utilized for In Vitro Fertilization (IVF), Artificial Insemination
(AI), Intrauterine Insemination (IUI), Gamete Intra-Fallopian
Transfer (GIFT), Zygote Intra-Fallopian Transfers (ZIFT), Intra-Cytoplasmic
Sperm Injection (ICSI), sex selection (shifting the sex ratio
via the use of an X or Y bearing sperm fraction) and other assisted
reproductive clinical procedures.
* * * * *
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