JP7092886B2 - 鳥類の卵から分離した細胞である胚盤葉多能性幹細胞、神経幹細胞、胚性線維芽細胞条件培地を有効成分とする細胞の増殖、肌の再生、及びシワ改善用の培地組成物 - Google Patents
鳥類の卵から分離した細胞である胚盤葉多能性幹細胞、神経幹細胞、胚性線維芽細胞条件培地を有効成分とする細胞の増殖、肌の再生、及びシワ改善用の培地組成物 Download PDFInfo
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Description
<1-1>鳥類の卵からの神経幹細胞の分離及び培養
赤色野鶏の1種類である白色レグホン種(White Leghorn)種の卵(egg)を用いて、神経幹細胞を分離及び培養した。
本発明の多能性幹細胞とは、受精卵の胚盤葉細胞から分離・培養した胚性幹細胞(Embryonic stem cell;ESC)を意味し、OCT4、NANOG、SOX2、アルカリフォスファターゼ(Alkaline phosphatase)活性を有している細胞を意味する。
赤色野鶏の1種類である白色レグホン種(White Leghorn)種の卵(egg)を用いた。卵受精卵を4日間37℃~38℃の条件下で培養して得た組織を取った後、頭と内臓を除去してから、酵素(トリプシン-EDTA)で処理した。105%FBS入りDMEMを添加した後、800gで3分間遠心分離した後、上澄み液を除去し、10%DMEM培地で長期にわたって培養して、胚性線維芽細胞(embryonic fibroblasts;FBs)を得た。具体的な培養方法については、Dai R et al(2014).Stem cell international 2014:1-15を参照した。
前記<実施例1>において得た卵由来の幹細胞である多能性幹細胞、または神経幹細胞、及び線維芽細胞を用いて条件培地を製造した。
前記<実施例1>において分離及び培養した卵由来の幹細胞の幹細胞能を確認した。多能性幹細胞は、アルカリフォスファターゼ(Alkaline phosphatase)活性及びマーカー遺伝子であるOCT4、SOX2、NANOG遺伝子を発現させ、神経幹細胞は、アルカリフォスファターゼ活性及びマーカー遺伝子であるNestin、SOX9を発現させるため、アルカリフォスファターゼ染色、RT-PCRまたは免疫化学染色を用いて卵由来の幹細胞の幹細胞能を確認した。
鳥類の卵幹細胞条件培地のヒト幹細胞の増殖能を確認するために、血清代替物(Serum Replacer)(20%)入りDMEM/F12培地(SR)と前記<実施例2>において製造した卵由来の多能性幹細胞条件培地(PCS-CM)上でヒト胚性幹細胞または間葉系幹細胞をそれぞれ培養した後、結果を比較・分析した。
鳥類の卵幹細胞条件培地の皮膚細胞の増殖能を確認するために、前記<実施例1>において分離した線維芽細胞、神経幹細胞、多能性幹細胞条件培地を4%の濃度で含んでいる基本培地で皮膚細胞を培養した。
鳥類の卵幹細胞条件培地の肌の再生及びシワ改善の効果を確認した。
Claims (9)
- ニワトリの卵(Egg)由来の幹細胞条件培地(conditioned medium)を有効成分として含む細胞増殖用の培地組成物であって、
前記ニワトリの卵由来の幹細胞は、卵細胞から抽出した多能性幹細胞(pluripotent stem cells;PSCs)または神経幹細胞であり、
前記細胞増殖用は、ヒト幹細胞または皮膚細胞増殖用である、細胞増殖用の培地組成物。 - 前記ヒト幹細胞は、胚性幹細胞または成体幹細胞であることを特徴とする、請求項1に記載の細胞増殖用の培地組成物。
- 前記成体幹細胞は、骨髄由来、臍帯血由来、脂肪由来、血液由来、肝臓由来、皮膚由来、胃腸管由来、胎盤由来、神経由来、副腎由来、上皮由来及び子宮由来の幹細胞からなる群より選ばれるいずれか1種であることを特徴とする、請求項2に記載の細胞増殖用の培地組成物。
- ニワトリの卵(Egg)由来の細胞条件培地を有効成分として含み、
前記ニワトリの卵由来の細胞は、ニワトリの卵細胞から抽出した多能性幹細胞、神経幹細胞及び線維芽細胞からなる群より選ばれるいずれか1種である、創傷治療用の薬学的組成物。 - ニワトリの卵(egg)由来の細胞条件培地を有効成分として含有し、
前記ニワトリの卵由来の細胞は、ニワトリの卵細胞から抽出した多能性幹細胞、神経幹細胞及び線維芽細胞からなる群より選ばれるいずれか1種である、肌の再生またはシワ改善用の化粧料組成物。 - 前記ニワトリの卵(egg)由来条件培地は、コラーゲン遺伝子の発現を増加させ、ヒアルロン酸の生成を増加させることを特徴とする、請求項5に記載の肌の再生またはシワ改善用の化粧料組成物。
- 前記ニワトリの卵(egg)由来条件培地は、エラスチン遺伝子の発現を増加させることを特徴とする、請求項5に記載の肌の再生またはシワ改善用の化粧料組成物。
- 前記組成物は、化粧水、皮膚軟化剤、スキントナー、アストリンジェント、ローション、乳液、モイスチャーローション、栄養ローション、マッサージクリーム、栄養クリーム、モイスチャークリーム、ハンドクリーム、ファンデーション、エッセンス、栄養エッセンス、パック、石鹸、クレンジングフォーム、クレンジングローション、クレンジングクリーム、ボディローション、ボディクレンザー、洗顔剤、トリートメント、美容液、美容パック、軟膏剤、ゲル剤、塗布剤、液剤、パッチ及び噴霧剤からなる群より選ばれるいずれか1種に剤形化されることを特徴とする、請求項5に記載の肌の再生またはシワ改善用の化粧料組成物。
- ニワトリの卵(Egg)由来の幹細胞条件培地を細胞と培養するステップを含み、
前記ニワトリの卵由来の幹細胞は、ニワトリの卵細胞から抽出した多能性幹細胞(pluripotent stem cells;PSCs)または神経幹細胞であり、
前記細胞は、ヒト幹細胞または皮膚細胞である、細胞の増殖方法。
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KR1020170166745A KR102121042B1 (ko) | 2017-12-06 | 2017-12-06 | 조류의 에그로부터 분리한 세포인 배반엽 전분화능세포, 신경줄기세포, 배아섬유아세포 컨디션드 배지를 유효성분으로 하는 세포 증식, 피부재생 및 주름 개선용 배지 조성물 |
PCT/KR2018/011852 WO2019112161A2 (ko) | 2017-12-06 | 2018-10-10 | 조류의 에그로부터 분리한 세포인 배반엽 전분화능세포, 신경줄기세포, 배아섬유아세포 컨디션드 배지를 유효성분으로 하는 세포 증식, 피부재생 및 주름 개선용 배지 조성물 |
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CN106318904B (zh) * | 2015-06-15 | 2020-08-11 | 上海朗斯生物工程有限公司 | 间充质干细胞条件培养基用于美容品领域的用途 |
CN104997708A (zh) * | 2015-07-07 | 2015-10-28 | 河南中科干细胞基因工程有限公司 | 干细胞条件培养基的获取及在防皮肤老化方面的应用 |
KR20180007256A (ko) * | 2016-07-12 | 2018-01-22 | 이정복 | 에그 세포 배양액을 유효성분으로 포함하는 인간 줄기세포 성장용 배지 조성물 |
CN106511387A (zh) * | 2016-10-26 | 2017-03-22 | 谢培增 | 鸡胚胎素制备方法及应用 |
KR101830062B1 (ko) * | 2017-06-30 | 2018-02-20 | 주식회사 차바이오텍 | 피부줄기세포 배양액을 포함하는 피부 노화 예방 또는 개선용 조성물 및 이의 용도 |
KR101849631B1 (ko) * | 2018-03-26 | 2018-04-17 | 이정복 | 에그 세포 배양액을 유효성분으로 포함하는 인간 줄기세포 성장용 배지 조성물 |
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2017
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JP2010524482A (ja) | 2007-04-24 | 2010-07-22 | ビバリス | ウイルスワクチンの生産用のアヒル胚由来幹細胞株 |
JP2011505853A (ja) | 2007-12-17 | 2011-03-03 | アンスティテュ ナショナール ド ラ ルシェルシュ アグロノミク | 幹細胞および体細胞を胚芽細胞にイン・ビトロで再プログラムするための方法 |
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KR102121042B1 (ko) | 2020-06-09 |
WO2019112161A2 (ko) | 2019-06-13 |
WO2019112161A3 (ko) | 2019-07-25 |
US20200384036A1 (en) | 2020-12-10 |
EP3722414A2 (en) | 2020-10-14 |
EP3722414A4 (en) | 2021-10-06 |
KR20190066888A (ko) | 2019-06-14 |
JP2021505202A (ja) | 2021-02-18 |
CN111511901A (zh) | 2020-08-07 |
US11564951B2 (en) | 2023-01-31 |
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