JP5065885B2 - 前立腺幹細胞 - Google Patents
前立腺幹細胞 Download PDFInfo
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- JP5065885B2 JP5065885B2 JP2007503421A JP2007503421A JP5065885B2 JP 5065885 B2 JP5065885 B2 JP 5065885B2 JP 2007503421 A JP2007503421 A JP 2007503421A JP 2007503421 A JP2007503421 A JP 2007503421A JP 5065885 B2 JP5065885 B2 JP 5065885B2
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Description
i) 前立腺組織由来の前立腺細胞を含む細胞調製物を提供する工程、
ii) 上記前立腺細胞の培養状態での維持及び前立腺細胞のコラーゲン系マトリックスへの結合を可能にする細胞培養条件を提供する工程、
iii) CD133抗原を発現する結合細胞を選択する工程
を含む。
i) CD133抗原を発現する細胞を、顆粒球マクロファージコロニー刺激因子(GM−CSF)、幹細胞因子(SCF)及び白血病阻害因子(LIF)を含む培養培地中で培養する工程、及び
ii) 選択した(i)の細胞を血清無含有培地中で継代培養する工程
を含む。
i) 本発明に記載の少なくとも1つの前立腺幹細胞を含む調製物を提供する工程、
ii) 核酸を上記細胞調製物から抽出する工程、
iii) 上記抽出された核酸を核酸アレイと接触させる工程、及び
iv) 該核酸の該核酸アレイ上の結合パートナーへの結合を示すシグナルを検出する工程
を含む、前立腺幹細胞に関連した遺伝子の同定方法が提供される。
i) 上記核酸の上記結合パートナーへの結合によって生成されるシグナル(1つ又は複数)を照合する工程、
ii) 照合されたシグナル(1つ又は複数)をデータ解析可能な形態に変換する工程、及び
任意選択で、
iii) 解析されたデータの出力を提供する工程
を含む。
i) 本発明に記載の少なくとも1つの前立腺幹細胞を含む調製物を提供する工程、
ii) 核酸を上記細胞調製物から抽出する工程、
iii) 上記抽出された核酸に含まれるリボ核酸からcDNAを調製する工程、及び
iv) (iii)で形成したcDNAをベクター内にライゲートする工程
を含む、前立腺特異的遺伝子発現産物を含むライブラリーの作製方法が提供される。
i) a) 本発明に記載の前立腺幹細胞、
b) (a)の上記細胞が接着して増殖し得る細胞培養物支持マトリックス、
c) 血清、間質画分並びに或る比率でのホルモンとしてエストロゲン及びジヒドロテストステロン又はその機能的誘導体を加えた細胞培養培地
を含む細胞培養容器を提供する工程、及び
ii) 上記容器内での上記前立腺由来細胞の成長及び分化を促進する条件を提供する工程
を含む、前立腺様腺房の形成のためのインビトロ方法が提供される。
i) 血清、間質画分並びに或る比率でのエストロゲン及びジヒドロテストステロン又はその機能的誘導体を加えた細胞培養培地中で形成させた本発明による前立腺幹細胞から形成された前立腺様腺房、細胞培養物支持マトリックス、及び前立腺腺房の成長を支持する細胞培養培地を含む細胞培養容器を提供する工程、及び
ii) 好ましくは活性化された内皮細胞を上記細胞培養容器に添加する工程であって、内皮細胞が増殖及び/又は移動して、血管細管を上記腺房内又はその周辺に形成する、添加する工程
を含む、血管新生した前立腺腺房の形成のためのインビトロ方法が提供される。
i) a) 前立腺幹細胞及び活性化された内皮細胞、
b) (a)の上記細胞が接着して増殖し得る細胞培養物支持マトリックス、
c) 血清、間質画分並びに或る比率でのホルモンとしてエストロゲン及びジヒドロテストステロン又はその機能的誘導体を加えた細胞培養培地
を含む細胞培養容器を提供する工程、及び
ii) 前立腺腺房を形成させるための上記容器内での上記前立腺幹細胞の成長及び分化、並びに上記活性化された内皮細胞からの血管細管の形成による該腺房の血管新生を促進させる条件を提供する工程
を含む、血管新生した前立腺様腺房の形成のためのインビトロ方法が提供される。
i) 培養条件及び本発明に記載の少なくとも1つの癌性腺房を提供する工程、
ii) 少なくとも1種類の被験薬剤を添加する工程、及び
iii) 上記薬剤の抗増殖活性を、癌性腺房を含む細胞に関してモニターする工程
を含む、癌性前立腺細胞の増殖を阻害できる薬剤の同定方法が提供される。
i) 培養条件及び本発明に記載の少なくとも1つの癌性腺房を提供する工程、
ii) 少なくとも1種類の被験薬剤を添加する工程、及び
iii) 癌性腺房を含む細胞の運動性をモニターする工程
を含む、癌性前立腺細胞の運動性を阻害できる薬剤の同定方法が提供される。
i) 本発明に記載の前立腺幹細胞を含む調製物を提供する工程、及び
ii) 発現が前立腺細胞の分化と関連する少なくとも1つの遺伝子の発現を測定する工程
を含む、前立腺細胞分化のマーカーの同定方法が提供される。
i) 本発明に記載の前立腺幹細胞を含む調製物を提供する工程、及び
ii) 発現が前立腺細胞の形質転換と関連する少なくとも1つの遺伝子の発現を測定する工程
を含む、前立腺細胞の形質転換のマーカーの同定方法が提供される。
ハイブリダイゼーション: 5×SSCで65℃にて16時間
洗浄2回: 各々、2×SSCで室温(RT)にて15分間
洗浄2回: 各々、0.5×SSCで65℃にて20分間
ハイブリダイゼーション: 5×〜6×SSCで65℃〜70℃にて16〜20時間
洗浄2回: 各々、2×SSCでRTにて5〜20分間
洗浄2回: 各々、1×SSCで55℃〜70℃にて30分間
ハイブリダイゼーション: 6×SSCでRT〜55℃にて16〜20時間
少なくとも2回洗浄: 各々、2×〜3×SSCでRT〜55℃にて20〜30分間
i) 前立腺細胞を含む単離試料を提供する工程、及び
ii) 図3に示す核酸配列を含む核酸分子、又は核酸にストリンジェントハイブリダイゼーション条件下でハイブリダイズする核酸分子の発現を検出する工程
を含む、被験体を前立腺癌又は前立腺癌に対する素因についてスクリーニングする方法が提供される。
i) 前立腺幹細胞の調製物を提供する工程、
ii) 上記細胞を非ヒト動物被験体内に移植する工程、及び
iii) 移植された細胞の分化及び成長をモニターする工程
を含む、前立腺腺房の形成の解析のための非ヒト動物モデルが提供される。
単離した腫瘍幹細胞の遺伝子型
散発性前立腺癌と関連するマイクロサテライトマーカー(8p 10q 16p、図3)の組合せを用い、単離したHEA+/CD44+/α2β1 hi/CD133+細胞が、同じ患者由来の血中リンパ球DNAと比べ、前立腺腫瘍に特徴的なヘテロ接合性パターンの減損を示すか否かを判定することができる。この解析は、3MM紙及び蛍光標識したPCRプライマーを用いた培養物のマイクロサンプリングにて行なう(MacIntosh他、1998)。これにより、正常細胞と癌細胞とを区別することができ、幹細胞が確かに形質転換事象の標的であるかを判定することができる。
性質が異なる腫瘍細胞集団を単離し、その増殖、分化及び悪性の可能性をインビトロ及びインビボで判定する。下記の集団(HEA+/CD44−(管腔細胞)、HEA+/CD44+(基底細胞)、HEA+/CD44+/α2β1 low/CD133−(移行(transit)細胞)、HEA+/CD44+/α2β1 hi/CD133+(幹細胞)を単離し、未分類腫瘍集団と比較する。
性質が異なる集団(上記)の癌細胞の形質転換能(足場非依存性)を、当該細胞の軟質寒天中でのコロニー形成能によって測定する。個々のコロニーを、21日後に、倒立顕微鏡を用いて計測する。CFEとコロニーの大きさの比較を行なう。
本発明者らは、腫瘍幹細胞及びその前駆体が再構成基底膜(例えば、Matrigel)中で腺の形態形成が行われる可能性を調べた。本発明者らにより、正常基底細胞が、コラーゲン系マトリックス(例えば、Matrigel)中で間質とともに、アンドロゲンの存在下で培養すると腺の形態形成が行われ得ることが示された。構造及び表現型がインビボ腺房と類似した球状形態が生成され、多くの場合、分岐した腺胞管様である(Lang他、2001)。対照的に、癌細胞は、多くの場合、紡錘形状の細胞の大きな集塊を形成するが、明白な器質化(organization)は見られない。それにもかかわらず、この構造体は、多くの場合、或る程度の分化を示す細胞を含有し、元の腫瘍との比較が可能である。
これらの幹細胞がMatrigel中を移動する能力を、改良ボイデン・チャンバ(Boyden-chamber)法(Albini他、1987)によって測定する。移動速度は、EGFPで標識した細胞を用い、経時的共焦点顕微鏡検査により評価する。本発明者らにより、低レベルのEGFPを発現する前立腺上皮が生成された。作製したpLNCX−EGFP(2)系組換えレトロウイルスを用いて細胞集団を感染させ、G418耐性コロニーを運動性アッセイに用いる。低レベルのGFP発現を用いて浸潤及び運動性をリアルタイムで追跡する。
腫瘍幹細胞は、正常幹細胞を規定する重要な基準を有するはずである。移植後、これらは、増殖、分化及び自己蘇生(self-renew)するはずである。異なる表現型の腫瘍のインビボでのコロニー形成能力を調べるため、幹細胞、移行細胞、基底細胞、管腔細胞及び未分類細胞の移植片を6〜8週齢の雄の免疫不全(immunocomprimised)マウスの前立腺内に導入する。マウスを、移植時に、徐放性テストステロンペレットの皮下移植によってホルモン処置する。各集団について、成功裡に生着し、且つ腫瘍増殖を開始する細胞の数を、移植する細胞の数を変えることにより測定する。性質が異なる集団の自己蘇生能力は、二次レシピエント内に連続移植することにより調べる。
発現プロフィールを、癌含有組織試料及び非癌組織試料から単離した幹細胞から得る。Affymetrix GeneChipマイクロアレイプラットフォームを用い、各試料について遺伝子発現の絶対レベルをアッセイする。これを行なうため、全RNAを精製α2β1 ++細胞から抽出する。細胞収率が低いため、アレイへのハイブリダイゼーションのために充分な標的を提供する線形増幅工程を用いることが必要である。Affymetrixの少量試料標識プロトコルでは、100ng(約104細胞)で効を奏する(work well)が、1〜10ngという少量の全RNAも使用可能であることが示された。これまで、本発明者らは、この技術(Hu−U133A GeneChips)を、本発明者らによって最近単離された前立腺癌リンパ節転移細胞株に由来するものである選択した細胞集団(α2β1 ++を含む)から抽出された、増幅した全RNAのプロフィールを調べるために使用している。
Claims (6)
- 以下の工程:
i) 前立腺組織由来の前立腺癌幹細胞を含む細胞調製物を提供する工程、
ii) 該前立腺癌幹細胞の培養状態での維持及び該前立腺癌幹細胞のコラーゲン系マトリックスへの結合を可能にする細胞培養条件を提供する工程、
iii) CD133抗原、CD44抗原、ヒト上皮抗原(HEA)、および高レベルのα2β1インテグリンを発現する前記結合された細胞を選択する工程
を含む、CD133抗原、CD44抗原、HEA、および高レベルのα2β1インテグリンを発現する前立腺癌幹細胞の選択的富化を含む、前立腺癌幹細胞の単離方法。 - 以下のさらなる工程:
i) CD133抗原、CD44抗原、HEA、および高レベルのα2β1インテグリンを発現する前立腺癌幹細胞を、顆粒球マクロファージコロニー刺激因子(GM−CSF)、幹細胞因子(SCF)及び白血病阻害因子(LIF)を含む培養培地中で培養する工程、及び
ii) (i)の前立腺癌幹細胞を血清無含有培地中で継代培養する工程
を含む、請求項1に記載の方法。 - 前記前立腺癌幹細胞が原発性で転移性の前立腺腫瘍に由来するものである、請求項1または2に記載の方法。
- 前記コラーゲン系マトリックスがI型コラーゲンを含む、請求項1〜3のいずれか1項に記載の方法。
- 高いインビトロ増殖能を有し、α2β1インテグリンlow/CD133−前立腺細胞よりも高いコロニー形成効率を有し、かつ、免疫不全非ヒト動物モデルにおいて癌性前立腺様腺房を形成することができる、請求項1〜4のいずれか1項に記載の方法によって得ることができる前立腺癌幹細胞。
- 高いインビトロ増殖能を有し、α2β1インテグリンlow/CD133−前立腺細胞よりも高いコロニー形成効率を有し、かつ、免疫不全非ヒト動物モデルにおいて癌性前立腺様腺房を形成することができる、CD133抗原、CD44抗原、HEA、および高レベルのα2β1インテグリンを発現する前立腺癌幹細胞の細胞培養物。
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US10538742B2 (en) | 2004-11-12 | 2020-01-21 | Cambridge Enterprise Limited | Methods and means related to cancer stem cells |
US20090012024A1 (en) * | 2005-07-26 | 2009-01-08 | Procure Therapeutics Limited | Stem Cell Markers |
US8647867B2 (en) | 2005-09-30 | 2014-02-11 | National University Corportion Okayama University | Cell cultivation method and cell culture |
GB0625041D0 (en) * | 2006-12-15 | 2007-01-24 | Procure Therapeutics Ltd | Prostate stem cell model |
HUE027617T2 (en) * | 2006-12-20 | 2016-11-28 | Novarx | Universal tumor cell vaccine for therapeutic and prophylactic use |
US8309354B2 (en) | 2007-01-22 | 2012-11-13 | Macrogenics West, Inc. | Human cancer stem cells |
GB0701321D0 (en) * | 2007-01-24 | 2007-03-07 | Procure Therapeutics Ltd | Agent |
GB0908934D0 (en) | 2009-05-26 | 2009-07-01 | Univ The West Of Scotland | Differentiation medium |
GB0910751D0 (en) | 2009-06-23 | 2009-08-05 | Procure Therapeutics Ltd | Prostate cancer vaccine |
WO2011158019A1 (en) | 2010-06-16 | 2011-12-22 | Adjuvantix Limited | Polypeptide vaccine |
WO2012010904A1 (en) | 2010-07-23 | 2012-01-26 | Procure Therapeutics Limited | Mammalian model for amplification of cancer stem cells |
WO2012026712A2 (ko) * | 2010-08-23 | 2012-03-01 | 주식회사 강스템홀딩스 | Nod2의 아고니스트를 처리한 줄기세포 또는 그 배양물을 포함하는 면역질환 또는 염증질환의 예방 또는 치료용 약학조성물 |
CA2809864A1 (en) | 2010-09-03 | 2012-03-08 | Stem Centrx, Inc. | Identification and enrichment of cell subpopulations |
US9778264B2 (en) | 2010-09-03 | 2017-10-03 | Abbvie Stemcentrx Llc | Identification and enrichment of cell subpopulations |
GB201116702D0 (en) | 2011-09-28 | 2011-11-09 | Procure Therapeutics Ltd | Cell surface markers |
US9885718B2 (en) | 2014-07-02 | 2018-02-06 | Dragon Victory Development Ltd. | Specific biomarker set for non-invasive diagnosis of liver cancer |
US9506925B2 (en) | 2014-07-02 | 2016-11-29 | Dragon Victory Development Ltd. | Specific biomarker set for non-invasive diagnosis of liver cancer |
AU2014399919B2 (en) * | 2014-07-02 | 2019-10-24 | Dragon Victory Development Ltd. | Specific biomarker set for non-invasive diagnosis of liver cancer |
Family Cites Families (17)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
DE68913658T3 (de) | 1988-11-11 | 2005-07-21 | Stratagene, La Jolla | Klonierung von Immunglobulin Sequenzen aus den variablen Domänen |
US6291158B1 (en) | 1989-05-16 | 2001-09-18 | Scripps Research Institute | Method for tapping the immunological repertoire |
JPH06509578A (ja) | 1991-07-26 | 1994-10-27 | ユニバーシティ・オブ・ロチェスター | 悪性細胞利用による癌治療法 |
US5631236A (en) | 1993-08-26 | 1997-05-20 | Baylor College Of Medicine | Gene therapy for solid tumors, using a DNA sequence encoding HSV-Tk or VZV-Tk |
WO1999014328A2 (en) | 1997-09-17 | 1999-03-25 | Genentech, Inc. | Secreted and transmembrane polypeptides and nucleic acids encoding the same |
US5856136A (en) | 1996-07-03 | 1999-01-05 | Incyte Pharmaceuticals, Inc. | Human stem cell antigens |
WO1998051805A1 (en) | 1997-05-15 | 1998-11-19 | Abbott Laboratories | Reagents and methods useful for detecting diseases of the prostate |
KR100272659B1 (ko) | 1997-06-28 | 2000-12-01 | 김영환 | 반도체 소자의 금속배선 선폭 측정방법 |
KR100490077B1 (ko) | 1999-10-29 | 2005-05-17 | 제넨테크, 인크. | 항-전립선 줄기세포 항원 (psca) 항체 조성물 및 그의사용 방법 |
US6541249B2 (en) * | 1999-12-22 | 2003-04-01 | Human Genome Sciences, Inc. | Immortalized human stromal cell lines |
US6632620B1 (en) * | 2000-06-22 | 2003-10-14 | Andrew N. Makarovskiy | Compositions for identification and isolation of stem cells |
US20050003527A1 (en) | 2001-08-09 | 2005-01-06 | Shona Lang | Cell culture method for obtaining prostate-like acini |
US20030119080A1 (en) | 2001-10-15 | 2003-06-26 | Mangano Joseph A. | Strategies for the identification and isolation of cancer stem cells and non-cancerous stem cells |
CA2469204A1 (en) | 2001-12-07 | 2003-06-19 | Regents Of The University Of Michigan | Prospective identification and characterization of breast cancer stem cells |
ITTO20020311A1 (it) * | 2002-04-10 | 2003-10-10 | Medestea Int Spa | Procedimento per la preparazione di cellule staminali da tessuto muscolare e tessuto adiposo umano e cellule staminali ottenibili mediante t |
JP2005528115A (ja) | 2002-05-31 | 2005-09-22 | ザ ボード オブ トラスティーズ オブ ザ リーランド スタンフォード ジュニア ユニバーシティ | 幹細胞および癌幹細胞を同定および単離する方法 |
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US8153421B2 (en) | 2012-04-10 |
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