JP2022044559A - サケの精巣からの高純度pdrnの抽出方法 - Google Patents
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Abstract
Description
本発明のPDRNの抽出方法において、前記サケ科魚類は、サケ亜科、カワヒメマス亜科及びシロマス亜科からなる群から選択される亜科に属する魚種であることが好ましく、前記サケ科魚類は、河口から川の上流10kmまでの河川サケであることが好ましい。
コイ、フナあるいは金魚は、年中12℃以下で飼育する場合、いつでも、受精卵が必要とする12~18時間以前にhCG500IU/kgあるいは500IU/個体を注射して、注射した翌日の夜明けから朝に自然に受精卵を得ることができるように雌と雄の生殖巣が成熟することを確認した。また、ウナギの場合、雌は、サケ脳下垂体抽出物(salmon pituitary extract)を注射して配偶子(卵)を性成熟させるが、雄の場合、hCGを500IU/kgの濃度で毎週注射して、3ヶ月で成熟した精子を得ることができ、雌の成熟した成熟卵との混合で受精卵を得ることができる。
第1ステップ:精液と未成熟な精巣部分とを分離
サケ科魚類の精巣から精液と未成熟な精巣部分とを分離する。図4のD部分から流れる精液を集めて原料として使用し、C部分を、D部位から精液を全て抜き取った後、所定の位置で切断する。
未成熟な精巣(図4のA、B、C)を磨砕し、下記の精漿液(seminal plasma)を用いて希釈する。磨砕の過程は、精巣をはさみで細かく切ると、精巣の精細胞などを包んでいる膜を剥ぎ取ることができるが、この膜を剥ぎ取った後、手袋をはめた手で優しく潰す方法を用いる。
hCGは、human chorionic gonadotropinの略字であって、ヒト絨毛性性腺刺激ホルモン25~200IU/gで10分~1時間処理して、未成熟な精巣の細胞を人為的に精子に性成熟させる。具体的には、前記潰された精巣に精漿液を入れ、希釈した後、インペラが装着された低速撹拌機を用いてゆっくり撹拌しながら、hCGを定量処理する。ただし、インペラの速度が速いと、細胞が破壊されてしまい、PDRNとして不純物が増加する可能性が高くなるので注意しなければならない。
hCG処理した精巣希釈液を1,000~10,000RCF(Relative Centrifugal Force)の速度で20分以上遠心分離して、沈殿した、活性を有し得る精子を回収する。
前記回収された精子から通常の方法でPDRN(PolyDeoxyRiboNucleotide)を抽出する。この抽出過程は、様々な方法が可能であるので、省略する。本発明の特徴は、PDRNの抽出方法ではなく、前記に記載したように、高純度のPDRNを抽出できる人工精液の製造にあるためである。
本発明では、雄サケから、精液が出るD部分(図4参照)を除いて未成熟状態の精巣を採取した。ホルモン(hCG)の未処理時に、AとB部位では精子の活性がほとんど現れず、Cの場合にも1%前後の低い活性を示し、成熟した精液では100%の精子活性を示した。しかし、hCG処理した場合、精巣のA部位は3%前後に向上するが、B部位は35%、C部位は89%の高い活性を示しており、全重量の45%に該当するC部位の精巣を用いて成熟した精子を作り出せば、不純物の少ない高純度のPDRNを作り出すことができるものと判断した(図4、表1)。
河川に上がってきた雄サケの精巣のC部位を緩衝溶液で希釈した後、hCGの処理濃度による精子の活性は、処理しなかった場合に2%と非常に低く、50IU/g精巣の処理濃度まで継続して精子活性が増加し、50~200IU/g精巣の処理濃度で82%以上の精子活性を示した(表6)。
本発明に用いられた精巣は、河川に上がってきた雄サケの精巣のC部位を10gずつ用い、比較のために、正常に採取された精液30mLを用いた(表8)。そして、50IU/g及び100IU/g精巣の濃度でhCG処理した場合、精巣全体と上層液80%以上を使用した結果とに区分して導出した。
Claims (7)
- 1)サケ科魚類の精巣から精液と未成熟な精巣部分とを分離するステップと、
2)前記未成熟な精巣部分を磨砕及び人工精漿液下で希釈するステップと、
3)前記精巣の細胞を人為的に精子に性成熟させるために、前記精巣の希釈液にhCG(human chorionic gonadotropin;ヒト絨毛性性腺刺激ホルモン)をg精巣当たり25~200IUで処理するステップと、
4)10分~2時間経過後、hCG処理した精巣の希釈液を遠心分離して、活性を有することができる精子を回収するステップと、
5)前記回収された精子からPDRN(Poly Deoxy Ribo Nucleotide)を抽出するステップと、を含む、サケの精巣からの高純度PDRNの抽出方法。 - 前記サケ科魚類は、サケ亜科、カワヒメマス亜科及びシロマス亜科からなる群から選択される亜科に属する魚種であることを特徴とする、請求項1に記載のPDRNの抽出方法。
- 前記サケ科魚類は、河口から川の上流10kmまでの河川サケであることを特徴とする、請求項1に記載のPDRNの抽出方法。
- 前記1)ステップの前記未成熟な精巣は、成熟の程度が異なる部分を区分して分離することによって、0.7%以上の精子活性度を示す、成熟度が相対的に高い部位であることを特徴とする、請求項1に記載のPDRNの抽出方法。
- 前記4)ステップの時間経過は10分~1時間であることを特徴とする、請求項1に記載のPDRNの抽出方法。
- 前記未成熟な精巣は、精原細胞又は精細胞を含むことを特徴とする、請求項1に記載のPDRNの抽出方法。
- 前記5)ステップの前に精子を含む懸濁液に淡水を入れ、精子が有したATPを消尽させてPDRNの純度を高めるステップをさらに含むことを特徴とする、請求項1に記載のPDRNの抽出方法。
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Citations (7)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
US4929542A (en) * | 1987-10-30 | 1990-05-29 | Cornell Research Foundation, Inc. | In vitro screening test for mutagenicity and genotoxicity during spermatogenesis |
JP2004196701A (ja) * | 2002-12-18 | 2004-07-15 | Nissan Chem Ind Ltd | 細胞死抑制剤 |
JP2005245394A (ja) * | 2004-03-08 | 2005-09-15 | Nissei Bio Kk | 魚類白子からの二本鎖dnaの抽出・精製方法 |
KR20140019609A (ko) * | 2012-08-06 | 2014-02-17 | (주)비알뷰티플레볼루션 | 항염 및 피부 재생 효과를 갖는 화장료 조성물 |
JP2015146760A (ja) * | 2014-02-06 | 2015-08-20 | 学校法人立命館 | タモロコ属魚類精子の受精能の評価方法 |
CN107099501A (zh) * | 2017-05-27 | 2017-08-29 | 华南农业大学 | 一种诱导幼龄灵长类动物获取性腺配子的方法和应用 |
CN110747194A (zh) * | 2019-11-28 | 2020-02-04 | 王超云 | 一种小分子多聚脱氧核糖核苷酸及其制备与应用 |
Family Cites Families (12)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
IT1187833B (it) | 1985-12-12 | 1987-12-23 | Farmigea Spa | Procedimento per l ottenimento di polidesossiribonucleotidi non informazionali sostanzialmente puri e dotati di attivita biologiche e prodotto relativo |
FR2605466B1 (fr) | 1986-10-15 | 1991-08-16 | Salmson Pompes | Dispositif de surveillance de moteur electrique |
FR2676926B1 (fr) * | 1991-05-30 | 1993-10-01 | Pierre Flecchia | Compositions pharmaceutiques a base de polydesoxyribonucleotides et leur procede de preparation. |
JP2976014B2 (ja) * | 1996-01-19 | 1999-11-10 | 農林水産省水産庁養殖研究所長 | 精巣精子を用いたウナギの人工受精法 |
IT1391866B1 (it) * | 2008-10-30 | 2012-01-27 | Mastelli S R L | Composizione iniettabile di polinucleotidi per il trattamento di patologie osteoarticolari. |
KR101739409B1 (ko) * | 2011-10-21 | 2017-05-25 | 주식회사 파마리서치프로덕트 | 어류 정소로부터 분리된 디엔에이 단편 혼합물을 포함하는 연골 재생용 조성물 |
CN107287186A (zh) | 2016-04-05 | 2017-10-24 | 达特珂贝怡股份有限公司 | 从鱼类的精液分离多脱氧核糖核苷酸的方法、由所述方法获得的多脱氧核糖核苷酸及其用途 |
KR20180066331A (ko) | 2016-12-07 | 2018-06-19 | 경기대학교 산학협력단 | 중공사형 비소흡착제 제조방법 및 중공사 담체 |
KR102031152B1 (ko) * | 2018-03-05 | 2019-10-15 | 한상철 | 연어 유래 폴리디옥시리보뉴클레오티드를 갖는 코어쉘 나노캡슐을 함유하는 피부개선 기능성 화장료 조성물 및 그의 제조방법 |
KR20190139633A (ko) | 2018-06-08 | 2019-12-18 | (주) 비앤에프솔루션 | 어류 정액 및 정소에서 피디알엔을 추출하는 방법 |
CN109349271A (zh) * | 2018-11-29 | 2019-02-19 | 江苏省农业科学院宿迁农科所 | 一种大鳞鲃精子保存液的使用方法 |
CN110786321A (zh) * | 2019-12-05 | 2020-02-14 | 广西壮族自治区水产科学研究院 | 一种金钱鱼精子冷冻保存方法 |
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Patent Citations (7)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
US4929542A (en) * | 1987-10-30 | 1990-05-29 | Cornell Research Foundation, Inc. | In vitro screening test for mutagenicity and genotoxicity during spermatogenesis |
JP2004196701A (ja) * | 2002-12-18 | 2004-07-15 | Nissan Chem Ind Ltd | 細胞死抑制剤 |
JP2005245394A (ja) * | 2004-03-08 | 2005-09-15 | Nissei Bio Kk | 魚類白子からの二本鎖dnaの抽出・精製方法 |
KR20140019609A (ko) * | 2012-08-06 | 2014-02-17 | (주)비알뷰티플레볼루션 | 항염 및 피부 재생 효과를 갖는 화장료 조성물 |
JP2015146760A (ja) * | 2014-02-06 | 2015-08-20 | 学校法人立命館 | タモロコ属魚類精子の受精能の評価方法 |
CN107099501A (zh) * | 2017-05-27 | 2017-08-29 | 华南农业大学 | 一种诱导幼龄灵长类动物获取性腺配子的方法和应用 |
CN110747194A (zh) * | 2019-11-28 | 2020-02-04 | 王超云 | 一种小分子多聚脱氧核糖核苷酸及其制备与应用 |
Non-Patent Citations (2)
Title |
---|
HIGAKI SHOGO, ET. AL: ""In vitro differentiation of fertile sperm from cryopreserved spermatogonia of the endangered endemi", SCIENTIFIC REPORTS, vol. 7, JPN6022032426, 17 February 2017 (2017-02-17), pages 1 - 14, ISSN: 0004846316 * |
三浦猛: "魚類精子形成の分子制御機構", 日本水産学会誌, vol. 62巻4号, JPN6022032425, 15 July 1996 (1996-07-15), pages 547 - 550, ISSN: 0004846317 * |
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