JP2004510423A5

JP2004510423A5 -

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Publication number: JP2004510423A5
Application number: JP2002531941A
Authority: JP
Filing date: 2001-01-24
Publication date: 2008-02-28

Description

【特許請求の範囲】
【請求項１】哺乳動物を産生する方法であって、該方法は、以下：
ａ．哺乳動物種の雄から精子細胞を収集する工程；
ｂ．カテーテルに協調性の粘性を有する培地を利用して、人工授精サンプルを確立する工程；
ｃ．該人工授精サンプルをカテーテルに配置する工程；
ｄ．該哺乳動物種の雌が適切に授精する時期を決定する工程；
ｅ．光学要素を、該哺乳動物種の雌に膣内挿入する工程；
ｆ．該カテーテルを、該哺乳動物種の雌に膣内挿入する工程；
ｇ．該光学要素および該カテーテルを、該哺乳動物種の雌の膣を介して導く工程；
ｈ．子宮卵管接合部を、該哺乳動物種の雌内で光学的に位置確認する工程；
ｉ．該カテーテルを、該哺乳動物種の雌内の該子宮卵管接合部の近傍に位置づけする工程；
ｊ．該人工授精サンプルの少なくとも一部を、該カテーテルから押し出す工程；
ｋ．該人工授精サンプルの少なくとも一部を、該哺乳動物種の雌内の子宮卵管接合部の近位に置く工程；
ｌ．該光学要素および該カテーテルを、該哺乳動物種の雌から引き出す工程；
ｍ．該哺乳動物種の雌の卵を受精させる工程；
ｎ．該受精した卵から子孫哺乳動物を産生する工程、
を包含する、方法。
【請求項２】前記配置する工程が、前記カテーテルに少数の精子を配置する工程を包含する、請求項１に記載の方法。
【請求項３】前記哺乳動物種の雌の卵を受精させる工程が、従来の子宮体人工授精プロセスに統計的に匹敵する成功レベルで該哺乳動物種の雌の卵を受精させる工程を包含する、請求項１〜２のいずれか１項に記載の方法。
【請求項４】請求項５８に記載の哺乳動物の産生方法であって、前記収集した精子細胞の少なくともいくらかを凍結する工程および該凍結した収集精子細胞の少なくともいくらかを解凍する工程をさらに包含する、方法。
【請求項５】前記確立する工程が、前記収集した精子細胞からの新鮮な精子を用いて人工授精サンプルを確立する工程をさらに包含する、請求項１〜４のいずれか１項に記載の方法。
【請求項６】前記確立する工程は、選別された性特異的人工授精サンプルを確立する工程をさらに包含し、前記配置する工程は、カテーテル内に該選別された性特異的人工授精サンプルを配置する工程をさらに包含し、前記押し出す工程は、該カテーテルから、該選別された性特異的人工授精サンプルの少なくとも一部を押し出す工程をさらに包含し、前記置く工程は、前記哺乳動物種の雌の子宮卵管接合部の近傍に、該選別された性特異的人工授精サンプルの少なくとも一部を置く工程をさらに包含する、請求項１〜５のいずれか１項に記載の方法。
【請求項７】授精標本において前記収集した精子細胞の特徴を一時的に実質的に保持する工程、前記哺乳動物種の雌の発情時期を決定する工程、およびおよそ該発情時期に人工授精サンプルを確立するために、該授精標本の特性を変更する工程、をさらに包含する、請求項１〜６のいずれか１項に記載の方法。
【請求項８】前記収集する工程が、哺乳動物種の雄から精巣上体精子細胞を収集する工程をさらに包含する、請求項１〜７のいずれか１項に記載の方法。
【請求項９】請求項１〜８のいずれか１項に記載の方法であって、前記カテーテルから押し出された前記人工授精サンプルの少なくとも一部を吸引して、吸引された授精サンプルを作製する工程をさらに包含する、方法。
【請求項１０】請求項１〜９のいずれか１項に記載の方法であって、前記収集された精子細胞から、授精する可能性の高そうな望ましい精子を選択する工程をさらに包含し、前記確立する工程は、該受精に望ましい精子を使用して、人工授精サンプルを確立する工程をさらに包含する、方法。
【請求項１１】前記哺乳動物種の雌内の子宮卵管接合部の近傍の表面下に前記カテーテルを挿入する工程をさらに包含し、前記押し出す工程は、該哺乳動物種の雌内の子宮卵管接合部の近傍の表面下で該カテーテルから前記人工授精サンプルの少なくとも一部を押し出す工程をさらに包含し、そして前記置く工程は、該哺乳動物種の雌内の該子宮卵管接合部の近傍の該表面下に該人工授精サンプルの少なくとも一部を置く工程をさらに包含する、請求項１〜１０のいずれか１項に記載の方法。
【請求項１２】カテーテルに前記人工授精サンプルを配置する工程が、前記カテーテルに少数の精子を配置する工程をさらに包含する、請求項１〜１２のいずれか１項に記載の方法。
【請求項１３】前記カテーテルに少数の精子を配置する工程が、以下：約１０，０００，０００未満の精子、約５，０００，０００個未満の精子、約２，０００，０００個未満の精子、約１，０００，０００個未満の精子、約５００，０００個未満の精子、および約１００，０００個未満の精子からなる群より選択される数の精子を配置する工程をさらに包含する、請求項１２に記載の方法。
【請求項１４】前記哺乳動物種の雌の卵を受精させる工程は、従来の子宮体人工授精プロセスに統計的に匹敵する成功レベルで該哺乳動物種の雌の卵を受精させる工程をさらに包含し、該成功レベルが、以下：少なくとも約７５％の成功率、少なくとも約６５％の成功率、少なくとも約６０％の成功率、少なくとも約５０％の成功率、少なくとも約４５％の成功率、およびその種についての従来の人工授精の少なくとも約９０％、からなる群より選択される、請求項１〜１３のいずれか１項に記載の方法。
【請求項１５】前記哺乳動物種の雌の卵を受精させる工程が、従来の子宮体人工授精プロセスに統計的に匹敵する成功レベルで該哺乳動物種の雌の卵を受精させる工程をさらに包含し、該成功レベルは、以下：
少なくとも約０．０５以上の統計的Ｐ値を有する従来の子宮体人工授精プロセスに統計的に匹敵する成功レベルで該哺乳動物種の雌の卵を受精させること、
少なくとも約０．０５以上の統計的Ｐ値を有する５００，０００，０００個のウマ精子サンプルを用いる従来の子宮体人工授精プロセスに統計的に匹敵する成功レベルで該哺乳動物種の雌の卵を受精させること、
少なくとも約８０％以上の統計的β値を有する従来の子宮体人工授精プロセスに統計的に匹敵する成功レベルで該哺乳動物種の雌の卵を受精させること、
少なくとも約０．０５以上の統計的Ｐ値および少なくとも約８０％以上の統計的β値を有する従来の子宮体人工授精プロセスに統計的に匹敵する成功レベルで該哺乳動物種の雌の卵を受精させること、
からなる群より決定される、請求項１〜１４のいずれか１項に記載の方法。
【請求項１６】前記哺乳動物種の雄から精子細胞を収集する工程が、ウシ、ウマ、またはブタからなる群より選択される哺乳動物種の雄から精子細胞を収集する工程をさらに包含する、請求項１〜１５のいずれか１項に記載の方法。
【請求項１７】人工授精サンプルを確立する工程が、スキムミルク媒体を使用する工程をさらに包含する、請求項１〜１６のいずれか１項に記載の方法。
【請求項１８】人工授精サンプルを確立する工程が、ＴＡＬＰ培地、ＨＥＰＥＳ緩衝化タイロード培地、およびアンドロヘップ培地からなる群から選択される媒体を使用する工程をさらに包含する、請求項１〜１７のいずれか１項に記載の方法。
【請求項１９】前記カテーテルに協調性の粘性を有する媒体を利用する工程が、前記哺乳動物の血液よりも高い粘性を有する人工受精サンプルを作製する媒体を利用する工程をさらに包含する、請求項１〜１８のいずれか１項に記載の方法。
【請求項２０】前記カテーテルに協調性の粘性を有する媒体を利用する工程が、生理食塩水よりも高い粘性を有する人工受精サンプルを作製する媒体を利用する工程をさらに包含する、請求項１〜１９のいずれか１項に記載の方法。
【請求項２１】請求項１〜２０のいずれか１項に記載の方法であって、前記カテーテルに協調性の粘性を有する媒体を利用する工程が、哺乳動物のほぼ平均体温において、それぞれ、約１００ｃｐより高い粘性を有する媒体；約３００ｃｐより高い粘性を有する媒体；約１０００ｃｐより高い粘性を有する媒体；約３０００ｃｐより高い粘性を有する媒体；および約６０００ｃｐより高い粘性を有する媒体からなる群から選択される媒体を利用する工程をさらに包含する、方法。
【請求項２２】請求項１〜２１のいずれか１項に記載の方法であって、前記人工授精サンプルを確立する工程は、子宮鏡に適合性の媒体を使用して、人工授精サンプルを確立する工程をさらに包含する、方法。
【請求項２３】前記精子細胞を凍結する工程をさらに包含する、請求項１〜２２のいずれか１項に記載の方法。
【請求項２４】請求項１〜２３のいずれか１項に記載の方法であって、前記授精のために子宮鏡を用いる授精に適合性の容量を決定する工程をさらに包含し、前記確立する工程は、該子宮鏡を用いる授精に適合性の容量で人工授精サンプルを確立する工程をさらに包含する、方法。
【請求項２５】請求項７に記載の方法であって、前記授精標本の特性を変更する工程を開始して、前記哺乳動物種の雌について適切に受精することが決定された時期付近で、発情期付近にある人工授精サンプルを確立する工程をさらに包含する、方法。
【請求項２６】請求項２５に記載の方法であって、前記授精標本の特性を変更する工程を開始して、前記哺乳動物種の雌について適切に受精することが決定された時期付近で、発情期付近にある人工授精サンプルを確立する工程が、以下：該哺乳動物種の雌について適切に受精すること決定された該時期の約２４時間以内、該哺乳動物種の雌について適切に受精すること決定された該時期の約１２時間以内、該哺乳動物種の雌について適切に受精すること決定された該時期の約８時間以内、該哺乳動物種の雌について適切に受精すること決定された該時期の約３時間以内、該哺乳動物種の雌について適切に受精すること決定された該時期の約１時間以内、からなる群から選択される時間内で、該授精標本の特性を変更する工程を開始する工程を包含する、方法。
【請求項２７】前記授精標本中の前記精子細胞の特性を一時的に実質的に保持する工程が、該精子細胞を凍結する工程を包含し、そしてここで、およそ前記発情時期に人工授精サンプルを確立するために、該授精標本の属性を変更する工程が、およそ該発情期に該授精標本を解凍する工程を包含する、請求項７に記載の哺乳動物を産生する方法。
【請求項２８】前記哺乳動物種の雌の発情時期を決定する工程が、該哺乳動物種の雌における発情を誘導する工程を包含する、請求項７に記載の方法。
【請求項２９】前記哺乳動物種の雌における発情を誘導する工程が、以下：
ａ．該哺乳動物種の雌を、プロゲスタゲンの処置に約１０日間毎日供する工程；および
ｂ．該哺乳動物種の雌を、プロゲスタゲンの処置に約１０日間毎日供する工程を達成するほぼ１０日目に、該哺乳動物種の雌を、クロプロステノールの処置に少なくとも１回供する工程、
を包含する、請求項２８に記載の方法。
【請求項３０】前記哺乳動物種の雌内の前記子宮卵管接合部の近傍に前記吸引された人工授精サンプルの少なくとも一部を置く工程が、該哺乳動物種の雌内の該子宮卵管接合部の陰窩内に該吸引された人工授精サンプルの少なくとも一部を置く工程を包含する、請求項９に記載の哺乳動物を産生する方法。
【請求項３１】請求項１〜３０のいずれか１項に記載の方法であって、前記哺乳動物種の雌内の子宮卵管接合部の近位の表面に対する前記精子の付着を促進させる工程をさらに包含する、方法。
【請求項３２】請求項３１に記載の方法であって、前記哺乳動物種の雌内の子宮卵管接合部の近位の表面に対する前記精子の付着を促進させる工程が、前記人工授精サンプルの少なくとも一部からフロスを作製する工程を包含する、方法。
【請求項３３】請求項１〜３２のいずれか１項に記載の方法であって、前記人工授精サンプルの少なくとも一部からフロスを作製する工程をさらに包含する、方法。
【請求項３４】前記人工授精サンプルを確立する工程が、前記精子細胞を希釈する工程をさらに包含する、請求項１〜３３のいずれか１項に記載の方法。
【請求項３５】前記精子細胞を希釈する工程が、２：１未満の比への該精子細胞の希釈、５：１未満の比への該精子細胞の希釈、１０：１未満の比への該精子細胞の希釈、スキムミルク媒体を用いた希釈、ＴＡＬＰ媒体を用いた希釈、ＨＥＰＥＳ緩衝タイロード媒体を用いた希釈、およびアンドロヘップ媒体を用いた希釈からなる群より選択される、該精子細胞を希釈する工程を包含する、請求項３４に記載の方法。
【請求項３６】前記人工授精サンプルを確立する工程が、ほぼ室温に前記授精標本を冷却する工程をさらに包含する、請求項１〜３５のいずれか１項に記載の方法。
【請求項３７】前記授精する可能性の高そうな望ましい精子を選択する工程が、前記哺乳動物種の雄から収集されたより運動性の精子を濃縮する工程を包含する、請求項１０に記載の方法。
【請求項３８】前記哺乳動物種の雄から収集されたより運動性の精子を濃縮する工程が、Ｐｅｒｃｏｌｌ勾配を通じて該精子を遠心分離する工程を包含する、請求項３７に記載の方法。
【請求項３９】前記Ｐｅｒｃｏｌｌ勾配を通じて前記精子を遠心分離する工程が、９０：４５％のＰｅｒｃｏｌｌ勾配を用いる工程を包含する、請求項３８に記載の方法。
【請求項４０】前記Ｐｅｒｃｏｌｌ勾配を通じて前記精子を遠心分離する工程が、以下：
ａ．約２００ｇで約５分間、該Ｐｅｒｃｏｌｌ勾配を通じて該精子を遠心分離する工程；および
ｂ．約８００ｇで約１０分間、該Ｐｅｒｃｏｌｌ勾配を通じて該精子を遠心分離する工程、
をさらに包含する、請求項３８または３９に記載の方法。
【請求項４１】前記哺乳動物種の雄から収集されたより運動性の精子を濃縮する工程が、該精子の濃度を出発濃度の約２倍未満に限定する工程を包含する、請求項３７に記載の方法。
【請求項４２】前記人工授精サンプルを確立する工程が、約３０μｌと約１５０μｌとの間、約５００μｌ未満、約２３０μｌ未満、および約１００μｌからなる群より選択される容量の授精標本を確立する工程をさらに包含する、請求項１〜４１のいずれか１項に記載の方法。
【請求項４３】請求項１〜４２のいずれか１項に記載の方法であって、前記光学要素およびカテーテルを哺乳動物種の雌の膣を通して導く工程が、以下：
ａ．該光学要素および該カテーテルを、該哺乳動物種の雌の頸部を通して導く工程；
ｂ．該光学要素および該カテーテルを、該哺乳動物種の雌の管腔を通して導く工程；および
ｃ．該光学要素および該カテーテルを、該哺乳動物種の雌の子宮角を通して導く工程、
を包含する、方法。
【請求項４４】請求項４３に記載の方法であって、前記光学要素およびカテーテルを哺乳動物種の雌の頸部を通して導く工程が、該光学要素および該カテーテルを、該哺乳動物種の雌の頸部を通して手動で導く工程を包含し、前記光学要素およびカテーテルを哺乳動物種の雌の管腔を通して導く前記工程が、該光学要素および該カテーテルを、該哺乳動物種の雌の管腔を通して手動で導く工程を包含する、方法。
【請求項４５】請求項１〜４４のいずれか１項に記載の方法であって、照射要素を前記光学要素の付近に挿入する工程をさらに包含する、方法。
【請求項４６】請求項４５に記載の方法であって、前記照射要素をストロボする工程をさらに包含する、方法。
【請求項４７】請求項４３に記載の方法であって、前記光学要素およびカテーテルを哺乳動物種の雌の子宮角を通して導く工程が、該光学要素および該カテーテルを、該哺乳動物種の雌の子宮角を通して、光学的に導く工程を包含する、方法。
【請求項４８】請求項１〜４７のいずれか１項に記載の方法であって、前記哺乳動物種の雌における優性な卵胞を決定する工程をさらに包含する、方法。
【請求項４９】請求項４８に記載の方法であって、少なくとも約３５ｍｍのサイズの排卵前の卵胞を観察する工程をさらに包含する、方法。
【請求項５０】請求項１〜４９のいずれか１項に記載の方法であって、ここで、前記カテーテルを哺乳動物種の雌内の子宮卵管接合部の近傍に位置づけする工程が、以下：
ａ．該子宮卵管接合部の約３〜５ｃｍ以内に該カテーテルを配置する工程；次いで
ｂ．該カテーテルを該子宮卵管接合部に接触させる工程、を包含する、方法。
【請求項５１】請求項１〜５０のいずれか１項に記載の方法であって、前記哺乳動物種の雌の子宮に濾過空気を導く工程をさらに包含する、方法。
【請求項５２】請求項１〜５１のいずれか１項に記載の方法であって、前記哺乳動物種の雌の子宮から空気を抜く工程をさらに包含する、方法。
【請求項５３】添付の図面を参照して本明細書で実質的に記載される哺乳動物を産生する方法。
【請求項５４】添付の図面を参照して本明細書で実質的に記載される方法によって産生される哺乳動物。
【請求項５５】添付の図面を参照して本明細書で実質的に記載されるプロセスを利用して産生される動物。 [Claim of claim]
    1. A mammalTheA method of producing, said method comprising
  a. Collecting sperm cells from a male of mammalian species;
  b.Using a medium with a coordinated viscosity on the catheter,Establishing an artificial insemination sample;
  c. The artificial insemination sample to the catheterDeployProcess;
  d. Determining when the female of said mammalian species will properly inseminate;
  e. Intravaginal insertion of an optical element into a female of said mammalian species;
  f. Vaginally inserting the catheter into a female of the mammalian species;
  g. Guiding the optical element and the catheter through the female vagina of the mammalian species;
  h. Optically locating the oviductal junction within the female of the mammalian species;
  i. Positioning the catheter near the oviductal junction in a female of the mammalian species;
  j. Extruding at least a portion of the artificial insemination sample from the catheter;
  k.Placing at least a portion of the artificial insemination sample proximal to a uterine tubal junction in a female of the mammalian species;
  l. Withdrawing the optical element and the catheter from a female of the mammalian species;
  m. Female egg of the mammal speciesReceivedRefining process;
  n. Producing offspring mammals from said fertilized eggs;
Method, including
    [Claim 2] SaidThe process of arrangingSaidPlacing a small number of sperm on the catheter1Described inWhoLaw.
    3. The step of fertilizing female eggs of said mammalian species comprises the step of fertilizing female eggs of said mammalian species at a success level that is statistically comparable to a conventional endometrial artificial insemination process. , Claim 1Any one of ~ 2Described inWhoLaw.
    4. A method of producing a mammal according to claim 58, said method comprisingCollectedSperm cellsAt least some ofProcess of freezingAnd thawing at least some of said frozen harvested sperm cellsThe method further including
    [Claim 5] BeforeConfirmationThe standing process isFrom the collected sperm cellsTo establish artificial insemination sample using fresh spermfurtherClaims to be includedAny one of 1 to 4Described inWhoLaw.
    [6] The establishing step is selectedEstablishing a sex-specific artificial insemination sampleFurther comprising the placing step comprisingPlacing the sorted sex-specific artificial insemination sample in a catheterAnd the step of extrudingExtruding at least a portion of the sorted sex-specific artificial insemination sample from the catheterFurther comprising the step of placingPlacing at least a portion of the sorted sex-specific artificial insemination sample proximate a female uterotubal junction of a mammalian speciesThe method according to any one of claims 1 to 5, further comprising
    [Claim 7] In the insemination sampleSaid collectedTemporarily retaining characteristics of sperm cells substantially, SaidDetermining the estrus time of a female of mammalian species,andAltering the characteristics of the insemination sample to establish an artificial insemination sample approximately at the estrus timeThe method according to any one of claims 1 to 6, further comprising
    [Claim 8] BeforeRetentionCollecting the steps of collecting epididymal sperm cells from a mammal of male speciesfurtherIncludeThe method according to any one of claims 1 to 7Method.
    9. ClaimAny one of 1 to 8Described inWhoA method, further comprising aspirating at least a portion of the artificial insemination sample extruded from the catheter to produce an aspirated insemination sample.
    10. The claim of the inventionAny one of 1 to 9Described inWhoA method likely to inseminate the collected sperm cellsdesirableFurther include the step of selecting spermAnd the establishing step further includes establishing an artificial insemination sample using the sperm desired for the fertilization.how to.
    [Claim 11] BeforeMemorySubsurface near the oviductal junction in females of mammalian speciesSaidProcess of inserting a catheterfurtherIncludeThe step of extruding isExtruding at least a portion of the artificial insemination sample from the catheter below the surface near the oviductal junction in a female of the mammalian speciesFurther include, AndThe step of placing isPlacing at least a portion of the artificial insemination sample below the surface in the vicinity of the uterotubal junction in a female of the mammalian speciesfurtherClaims to be includedAny one of 1 to 10Described inWhoLaw.
    12. The step of disposing the artificial insemination sample on a catheter comprises:SaidPlace a few sperm on the catheterfurtherClaims to be includedAny one of 1 to 12Described inWhoLaw.
    13. The process of disposing a small number of sperms in the catheter includes the following: less than about 10,000,000 spermatozoa, less than about 5,000,000 spermatozoa, less than about 2,000,000 spermatozoa Placing a number of sperm selected from the group consisting of less than about 1,000,000 sperm, less than about 500,000 sperm, and less than about 100,000 spermfurtherClaims to be included12Described inWhoLaw.
    14. The step of fertilizing female eggs of said mammalian species comprisesAt a success level that is statistically comparable to the conventional uterine body artificial insemination processSaidFertilizing a female egg of a mammalian speciesFurther include the success levelBut with: at least about 75% success rate, at least about 65% success rate, at least about 60% success rate, at least about 50% success rate, at least about 45% success rate, and conventionally for that species Selected from the group consisting of at least about 90% of artificial insemination with,ClaimAny one of 1 to 13Described inWhoLaw.
    15. The aboveMammalian female eggsThe process of fertilization is at a level of success that is statistically comparable to conventional uterine body artificial insemination processesSaidA step of fertilizing a female egg of a mammalian speciesfurtherInclusion, the success level is:
  Fertilizing female eggs of said mammalian species at a success level that is statistically comparable to a conventional uterine body artificial insemination process having a statistical P value of at least about 0.05 or greater;
  A female egg of the mammalian species at a success level that is statistically comparable to a conventional uterine artificial artificial insemination process using 500,000,000 equine sperm samples having a statistical P value of at least about 0.05 or more. Fertilizing,
  Fertilizing female eggs of said mammalian species at a success level that is statistically comparable to a conventional uterine body artificial insemination process having a statistical β value of at least about 80% or higher;
  Fertilizing the female egg of said mammalian species at a success level that is statistically comparable to a conventional uterine body artificial insemination process having a statistical P value of at least about 0.05 or more and a statistical β value of at least about 80% or more To let
Claim determined from the group consisting ofAny one of 1 to 14Described inWhoLaw.
    16. Male to spermatozoa of said mammalian speciescellCollecting sperm from males of a mammalian species selected from the group consisting of cattle, horses, or pigscellProcess of collectingfurtherClaims to be includedAny one of 1 to 15Described inWhoLaw.
    [17] ManThe process of establishing the insemination sample isskim milkProcess of using mediafurtherClaims to be includedAny one of 1 to 16Described inWhoLaw.
    [18] ManEstablishing the engineering insemination sample using a medium selected from the group consisting of TALP medium, HEPES buffered Tyrodes medium, and Andro-Hep mediumfurtherClaims to be includedAny one of 1 to 17Described inWhoLaw.
    19. The catheterCooperative toUsing a medium having viscosity, using a medium producing an artificial insemination sample having a higher viscosity than the blood of said mammalfurtherClaims to be includedAny one of 1 to 18Described inWhoLaw.
    20. The catheterCooperative toThe process of utilizing a medium having viscosity further includes the process of utilizing a medium producing an artificial insemination sample having a viscosity higher than saline.Any one of 1 to 19Described inWhoLaw.
    Claim 21.Any one of 1 to 20Described inWhoAnd the catheterCooperative toA step of utilizing a medium having viscosity is, respectively, a medium having a viscosity higher than about 100 cp; a medium having a viscosity higher than about 300 cp; a medium having a viscosity higher than about 1000 cp; Utilizing a medium selected from the group consisting of a medium having a high viscosity; and a medium having a viscosity greater than about 6000 cp.furtherMethod of inclusion.
    Claim 22.Any one of 1 to 21Described inWhoThe lawThe step of establishing the artificial insemination sample comprisesUsing a medium compatible with hysteroscope,ManProcess of establishing a technical insemination samplefurtherMethod of inclusion.
    23. The method of claim 1, further comprising freezing the sperm cells.Any one of 1 to 22Described inWhoLaw.
    Claim 24.Any one of 1 to 23Described inWhoLaw and said inseminationforCapacity compatible with insemination using hysteroscopeFurther comprising the step of: determining, wherein said establishing step comprises a volume compatible with insemination using said hysteroscopesoArtificialinseminationsampleEstablishRukoAboutFurtherMethod of inclusion.
    Claim 25.7Described inWhoAct to initiate the step of altering the characteristics of the insemination sample to establish an artificial insemination sample in the vicinity of the estrus, around the time it was determined to properly fertilize the female of the mammalian species A method further comprising the steps.
    Claim 26.5Described inWhoAct to initiate the step of altering the characteristics of the insemination sample to establish an artificial insemination sample in the vicinity of the estrus, around the time it was determined to properly fertilize the female of the mammalian species Within about 24 hours of the time determined to properly fertilize the female of the mammalian species, and within about 12 hours of the time determined to properly fertilize the female of the mammalian species , Within about 8 hours of the time determined to properly fertilize the female of the mammalian species, within about 3 hours of the time determined to properly fertilize the female of the mammalian species Initiating the step of altering the property of the insemination sample within a time selected from the group consisting of within about one hour of the time determined to be properly fertilized for females of the species.
    27. The process of temporarily substantially retaining the properties of said sperm cells in said insemination sample comprises freezing said sperm cells, and whereinaboutSaid departureIn the seasonEstablish artificial insemination sampleForAnd changing the attributes of the insemination sample,aboutThe estrusIn the periodClaim: Including the step of thawing said insemination sample7A method of producing a mammal according to
    28. The method of determining the estrus time of a female of said mammalian species comprises the step of inducing estrus in a female of said mammalian species.7Described inWhoLaw.
    29. The process of inducing estrus in a female of said mammalian species comprises the steps of:
  a. Subjecting the female of said mammalian species daily to treatment of progestagen for about 10 days;
  b. Subjecting the female of the mammalian species to treatment of cloprostenol at least once approximately on day 10 to achieve subjecting the female of the mammalian species to treatment of progestagen daily for about 10 days;
Claims that include28Described inWhoLaw.
    30. The method according to claim 30, further comprising the steps of:Was aspiratedPlacing at least a portion of the artificial insemination sample into the crypts of the uterotubal junction in the female of the mammalian speciesWas aspiratedClaiming including placing at least a portion of the artificial insemination sample9A method of producing a mammal according to
    Claim 31.Any one of 1 to 30Described inWhoThe law, beforeMemoryMilk animalsseedPromoting the adhesion of said sperm to the proximal surface of the uterotubal junction within the female.
    Claim 32.31Described inWhoThe law, beforeMemoryMilk animalsseedPromoting adhesion of said spermatozoa to the proximal surface of the uterotubal junction in a female of the subject comprises producing floss from at least a portion of said artificial insemination sample.
    Claim 33.Any one of 1 to 32Described inWhoA method, further comprising the step of making a floss from at least a portion of the artificial insemination sample.
    34.Artificial insemination sampleEstablishing the step of diluting the sperm cells.furtherClaims to be includedAny one of 1 to 33Described inWhoLaw.
    35. The method of claim 1, wherein said step of diluting sperm cells comprises diluting the sperm cells to a ratio of less than 2: 1, diluting the sperm cells to a ratio of less than 5: 1, to a ratio of less than 10: 1. The sperm cells are selected from the group consisting of dilution of the sperm cells, dilution with skimmed milk medium, dilution with TALP medium, dilution with HEPES buffered Tyrodes medium, and dilution with Androhep medium Claim to include the step of diluting34Described inWhoLaw.
    36.Artificial insemination sampleThe process of establishing theSaidClaim further comprising the step of cooling the insemination sampleAny one of 1 to 35Described inWhoLaw.
    [37] SaidinseminationLikely toThe process of selecting the desired sperm isMemoryMilk speciesofClaiming includes the step of concentrating more motile sperm collected from males.10Described inWhoLaw.
    [38] BeforeMemoryMilk speciesMale ofClaim 30. The step of concentrating the more motile sperm collected from the step comprises centrifuging the sperm through a Percoll gradient.37Described inWhoLaw.
    39. The step of centrifuging the sperm through the Percoll gradient comprises using a 90: 45% Percoll gradient.38Described inWhoLaw.
    40. The step of centrifuging the sperm through the Percoll gradient comprises the steps of:
  a. Centrifuging the sperm through the Percoll gradient at about 200 g for about 5 minutes; and
  b. Centrifuging the sperm through the Percoll gradient at about 800 g for about 10 minutes;
Claim further including38 or 39Described inWhoLaw.
    [41] BeforeMemoryMilk speciesofThe process of concentrating more motile sperm collected from a male comprises limiting the concentration of said sperm to less than about twice the starting concentration.3Described in 7WhoLaw.
    42.Artificial insemination sampleThe step of establishing is selected from the group consisting of between about 30 μl and about 150 μl, less than about 500 μl, less than about 230 μl, and about 100 μlVolumeProcess to establish the amount of insemination samplefurtherClaims to be includedAny one of 1 to 41Described inWhoLaw.
    Claim 43.Any one of 1 to 42Described inWhoA method of directing the optical element and the catheter through the vagina of a female of mammalian species is:
  a. Guiding the optical element and the catheter through the female neck of the mammalian species;
  b. Directing the optical element and the catheter through the female lumen of the mammalian species;
  c. Guiding the optical element and the catheter through the female uterine horn of the mammalian species,
Method, including
    Claim 44.43Described inWhoThe method wherein directing the optical element and the catheter through a female neck of a mammalian species comprises manually guiding the optical element and the catheter through a female neck of the mammalian species, The step of directing the optical element and the catheter through a female lumen of a mammalian species comprises manually directing the optical element and the catheter through a female lumen of the mammalian species.
    45. The claim of the inventionAny one of 1 to 44Described inWhoA method, further comprising the step of inserting an illumination element in the vicinity of said optical element.
    Claim 46.45Described inWhoA method, further comprising strobing the illumination element.
    Claim 47.43Described inWhoThe method wherein directing the optical element and the catheter through the uterine horn of a mammalian species comprises optically guiding the optical element and the catheter through the uterine horn of the female species of the mammalian species how to.
    Claim 48.Any one of 1 to 47Described inWhoA method, further comprising the step of determining the dominant follicle in females of said mammalian species.
    Claim 49.48Described inWhoThe method further comprising the step of observing the preovulatory follicle at least about 35 mm in size.
    Claim 50.Any one of 1 to 49Described inWhoThe method, wherein the step of positioning the catheter near the oviductal junction in a female of mammalian species comprises:
  a. Placing the catheter within about 3-5 cm of the uterotubal junction; then
  b. Contacting the catheter with the oviductal junction.
    Claim 51.Any one of 1 to 50Described inWhoA method, further comprising the step of directing filtered air to the female uterus of said mammalian species.
    Claim 52.Any one of 1 to 51Described inWhoA method, further comprising the step of venting the female uterus of said mammalian species.
53. A method of producing a mammal substantially as described herein with reference to the accompanying drawings.
[54] A mammal produced by the method substantially as herein described with reference to the accompanying drawings.
55. The present specification substantially refers to the attached drawings.An animal produced using the process described in

Claims

A method of producing a mammal, said method comprising the steps of:
Collecting sperm cells from a male of mammalian species;
Establishing an artificial insemination sample from the collected sperm cells;
Placing the artificial insemination sample in a catheter;
Intravaginal insertion of an optical element into a female of said mammalian species;
Vaginally inserting the catheter into a female of the mammalian species;
Guiding the optical element and the catheter through the female vagina of the mammalian species;
Visually locating the oviductal junction within the female of the mammalian species;
Positioning the catheter in proximity to the oviductal junction in a female of the mammalian species;
Extruding at least a portion of the artificial insemination sample from the catheter;
Depositing at least a portion of the artificial insemination sample in proximity to the uterotubal junction in a female of the mammalian species;
Withdrawing the optical element and the catheter from a female of the mammalian species;
Fertilizing the female egg of said mammalian species; and
Producing offspring mammals from said fertilized eggs;
Method, including

Hysteroscopic insemination sample, which includes:
Reservoir elements;
A catheter system, wherein the reservoir element is responsive;
A hysteroscopically compatible amount of sperm contained within the reservoir element,
To prepare the insemination sample.

3. The hysteroscopically compatible amount of sperm contained in the reservoir element comprises a small number of sperm cells as compared to the number of sperm cells typically found in natural insemination. Insemination sample for hysteroscopy.

Compared to the number of sperm cells typically found in said natural insemination, a small number of sperm cells are:
Less than about 10 million sperm, less than about 5 million sperm, less than about 2 million sperm, less than about 1 million sperm, less than about 500,000 sperm, and less than about 100,000 sperm;
The hysteroscopic insemination sample according to claim 3, comprising a large number of sperm cells selected from the group consisting of

5. A hysteroscopic insemination sample according to claim 4, wherein the sperm cells comprise male-derived sperm cells of mammalian species selected from bovine, equine or porcine.

The hysteroscopically compatible amount of sperm contained within the reservoir element is:
Between about 30 μl and 150 μl, less than about 500 μl, less than about 230 μl, and less than about 100 μl,
The hysteroscopic insemination sample according to claim 2, comprising an amount selected from the group consisting of

The hysteroscopic insemination sample according to claim 2, wherein the hysteroscopically compatible amount of sperm contained in the reservoir element comprises sex-selected sperm cells from a male of mammalian species.

8. The hysteroscope of claim 7, wherein said male sex derived sperm cells of mammalian species comprise sperm cells of mammalian species selected from the group consisting of bovine, equine, or porcine. Insemination sample for examination.

A method of producing a mammal, said method comprising the steps of:
Collecting sperm cells from a male of mammalian species;
Freezing at least some of the collected sperm cells;
Thawing at least some of said frozen collected sperm cells;
Classifying the collected sperm cells according to sex specific characterization;
Establishing an artificial insemination sample from the collected sperm cells having a small number of spermatozoa compared to the natural insemination dose for the animal;
Placing the artificial insemination sample in a catheter;
Vaginally inserting the optical element into a female of said mammalian species;
Vaginally inserting the catheter into a female of said mammalian species;
Guiding the optical element and the catheter through the female vagina of the mammalian species;
Positioning the uterotubal junction within the female of the mammalian species within the vagina;
Positioning the catheter in proximity to a uterine tubal junction in a female of the mammalian species;
Extruding at least a portion of the artificial insemination sample from the catheter;
Depositing a small number of spermatozoa of said artificial insemination sample in the vicinity of said uterotubal junction in a female of said mammalian species;
Withdrawing the optical element and the catheter from a female of the mammalian species;
Fertilizing female eggs of said mammalian species at a success level that is statistically comparable to conventional uterine body artificial insemination processes;
Producing offspring animals from said fertilized eggs;
Method, including

10. A method of producing a mammal according to claim 9, wherein said step of collecting sperm cells from a male of said mammalian species comprises sperm from a male of mammalian species selected from the group consisting of cows, horses or pigs. Collecting the cells.

10. A method of producing a mammal according to claim 9, wherein the establishing step comprises establishing a classified sex-specific artificial insemination sample, and the positioning step comprises the classified sex-specific Placing the artificial artificial insemination sample in a catheter, the pushing out includes pushing out at least a portion of the classified sex specific artificial insemination sample out of the catheter, wherein the depositing comprises: Depositing at least a portion of the classified sex-specific artificial insemination sample proximate to the uterotubal junction within the female of mammalian species.

10. A method of producing a mammal according to claim 9, further comprising the step of determining when a female of said mammalian species is properly fertilized.

10. A method of producing a mammal according to claim 9, wherein the step of fertilizing female eggs of said mammal species comprises the statistics of a conventional uterine body artificial insemination process. Fertilizing female eggs of said mammalian species at a level of success that is scientifically comparable.

10. A method of producing a mammal according to claim 9, wherein said establishing step comprises the step of establishing an artificial insemination sample utilizing fresh spermatozoa derived from said collected sperm cells.

10. A method of producing a mammal according to claim 9, further comprising the step of selecting spermatozoa desired to achieve more fertilization from said collected sperm cells, said establishing step Establishing the artificial insemination sample using the sperm desired for said fertilization.

10. A method of producing a mammal according to claim 9, further comprising the step of inserting the catheter below the surface in the vicinity of the oviductal junction in a female of said mammalian species. And said extruding comprises extruding at least a portion of said artificial insemination sample from said catheter below the surface in the vicinity of the uterotubal junction in a female of said mammalian species, and Allowing the deposition step to deposit at least a portion of the artificial insemination sample below the surface in the vicinity of the uterotubal junction in a female of the mammalian species.

10. A method of producing a mammal according to claim 9, further comprising aspirating at least a portion of the artificial insemination sample extruded from the catheter to produce an aspirated insemination sample. .

10. A method of producing a mammal according to claim 9, further comprising the step of determining a hysteroscopic compatible volume for artificial insemination, wherein said establishing step comprises Establishing the artificial insemination in a hysteroscopic compatible volume.

10. A method of producing a mammal according to claim 9, said method comprising temporarily substantially retaining the properties of said collected sperm cells in an insemination specimen, said female of said mammalian species. Determining an oestrus age of the subject and altering the characteristics of the fertilized specimen to establish an artificial insemination sample at about the oestrus age.

A method of producing a mammal according to claim 9, comprising a minority The step of placing the spermatozoa in the catheter is as follows:
Less than about 10 million sperm, less than about 5 million sperm, less than about 2 million sperm, less than about 1 million sperm, less than about 500,000 sperm, and less than about 100,000 sperm;
Further comprising the step of placing a large number of sperms selected from the group consisting of

21. A method of producing a mammal according to claim 20, wherein the step of fertilizing fertilizes female eggs of the mammalian species at a success level that is statistically comparable to a conventional uterine artificial insemination process. And the success level comprises:
Fertilizing female eggs of said mammalian species at a success level that is statistically comparable to a conventional uterine artificial insemination process with a statistical P value of at least about 0.05 or more; Fertilizing female eggs at a success level that is statistically comparable to a conventional uterine artificial insemination process using 500 million horse sperm samples, with a statistical P value of at least about 0.05 or greater;
Fertilizing female eggs of said mammalian species at a statistical β value of at least about 80% or greater, at a success level that is statistically comparable to conventional artificial insemination processes of the uterine body;
A female egg of said mammalian species is statistically subjected to a conventional uterine artificial insemination process with both a statistical P value of at least about 0.05 and a statistical β value of at least 80%. Fertilizing at comparable success levels,
A method determined from the group consisting of

21. A method of producing a mammal according to claim 20, wherein the step of fertilizing comprises at least about 75% success rate of at least about 75% of female artificial eggs of said species of said species. Mammal at a success level selected from the group consisting of 65% success rate, at least about 60% success rate, at least about 50% success rate, at least about 45% success rate, and at least about 90% success rate Fertilizing the female eggs of the species at a success level that is statistically comparable to a conventional uterine artificial insemination process.

23. A method of producing a mammal according to claim 22, wherein said step of collecting sperm cells from a male of said mammalian species comprises sperm from a male of mammalian species selected from the group consisting of cows, horses or pigs. Collecting the cells.

10. A method of producing a mammal according to claim 9, further comprising the step of determining when females of said mammalian species are properly fertilized.

A hysteroscopic insemination sample produced according to the method of any one of claims 1-24.

25. A mammal produced according to the method of any one of claims 1-24.

An insemination sample comprising a plurality of sperm cells contained within a reservoir element of a catheter system, wherein the sample is configured to be positioned near a female uterine oviductal junction of a mammalian species, and the uterus An insemination sample deposited in the vicinity of the oviductal junction in a female of the mammalian species by visually locating the oviductal junction.

28. The insemination sample of claim 27, comprising a hysteroscopic compatible medium contained within the reservoir.

28. The insemination sample according to claim 27, comprising a hysteroscopically compatible volume of sperm contained in the reservoir.

28. The insemination sample of claim 27, comprising a plurality of freeze-thawed sperm cells.

28. The insemination sample according to claim 27, comprising a plurality of classified sperm cells classified according to sex specific characterization.

An insemination sample comprising a plurality of sperm cells, wherein the sample is configured to be positioned in proximity to a female uterotubal junction of a mammalian species and wherein the uterotubal junction is visually located An insemination sample deposited within the vicinity of the oviductal junction in a female of the mammalian species by confirmation.

33. The insemination sample according to claim 32, comprising a hysteroscopic compatible medium.

33. The insemination sample according to claim 32, comprising a hysteroscopically compatible volume of sperm.

33. The insemination sample according to claim 32, comprising a plurality of freeze-thawed sperm cells.

33. The insemination sample according to claim 32, comprising a plurality of classified sperm cells classified according to sex specific characterization.

The hysteroscopic insemination sample according to claim 2, wherein the sample is configured to be positioned in proximity to a female uterine tubal junction of a mammalian species, and wherein the uterotubal junction is An insemination sample deposited within the vicinity of the uterotubal junction within the female of the mammalian species by visually locating.