JP2022013636A - 類似間葉系幹細胞の製造方法及びこれにより製造された類似間葉系幹細胞 - Google Patents
類似間葉系幹細胞の製造方法及びこれにより製造された類似間葉系幹細胞 Download PDFInfo
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Abstract
Description
Claims (16)
- 類似間葉系幹細胞の製造方法であって、
(a)細胞株の確立後、70回以下の継代培養されたヒト多能性幹細胞を準備するステップと、
(b)前記ヒト多能性幹細胞を分化誘導した胚様体のうち、包嚢胚様体を選別するステップと、
(c)前記包嚢胚様体を細胞透過性3次元培養挿入体上にロードして類似間葉系幹細胞を分離するステップと、
(d)前記細胞透過性3次元培養挿入体を透過した細胞のうち、単層(monolayer)形態の細胞群集のみを分離するステップと、
(e)前記単層形態の細胞群集を縦と横それぞれ100μm~500μmの大きさに均一化し、類似間葉系幹細胞を培養するステップと、
を含み、
前記類似間葉系幹細胞は、抗炎症効能及び免疫抑制能を有し、CD90及びSOX2を95%以上発現する、類似間葉系幹細胞の製造方法。 - 前記細胞透過性3次元培養挿入体は、ナイロン、繊維、ポリエチレン、ポリプロピレン、グラフェン、チタン、銅、ニッケル、銀、金、白金のいずれか1つ以上からなる、請求項1に記載の類似間葉系幹細胞の製造方法。
- 請求項1又は請求項2に記載の製造方法により製造されたヒト多能性幹細胞由来類似間葉系幹細胞において、
前記類似間葉系幹細胞は、骨髄由来間葉系幹細胞に比べてMMP-1タンパク質及びHGFタンパク質を高発現し、骨髄由来間葉系幹細胞に比べてCD95を低発現し、CD90及びSOX2を95%以上発現することを特徴とする、類似間葉系幹細胞。 - 前記類似間葉系幹細胞は、骨髄由来間葉系幹細胞に比べて前記MMP-1タンパク質を15倍以上高発現し、HGFタンパク質を2倍以上高発現する、請求項3に記載の類似間葉系幹細胞。
- 前記類似間葉系幹細胞は、骨髄由来間葉系幹細胞に比べてCD95を15倍以上低発現する、請求項3又は請求項4に記載の類似間葉系幹細胞。
- 前記類似間葉系幹細胞は、骨髄由来間葉系幹細胞に比べて炎症調節遺伝子を2倍~100倍以上高発現し、
前記炎症調節遺伝子は、Gata3、Adora2a、Gps2、Psma1、Pbk、Lrfn5、Cdh5、Apoe、Foxf1、Tek、Cx3cl1、Ptger4、Acp5、Bcr、Socs5、及びMdkからなる群のいずれか1つ以上である、請求項3~請求項5のいずれか一項に記載の類似間葉系幹細胞。 - 前記類似間葉系幹細胞は、Gata3、Adora2a、及びGps2遺伝子を同時に発現する、請求項3~請求項6のいずれか一項に記載の類似間葉系幹細胞。
- 前記類似間葉系幹細胞は、骨髄由来間葉系幹細胞に比べて免疫抑制能遺伝子を2倍~100倍以上高発現し、
前記免疫抑制能遺伝子は、Gata3、Gps2、Psma1、Apoe、Foxf1、Tek、Cx3cl1、Ptger4、Bcr、Socs5及びMdkのいずれか1つ以上である、請求項3~請求項7のいずれか一項に記載の類似間葉系幹細胞。 - 前記類似間葉系幹細胞は、Gata3、Gps2及びPsma1遺伝子を同時に発現する、請求項3~請求項8のいずれか一項に記載の類似間葉系幹細胞。
- 請求項3~請求項9のいずれか一項に記載の類似間葉系幹細胞を含む治療剤組成物であって、
前記治療剤組成物は、細胞治療剤組成物、細胞遺伝子治療剤組成物、組織工学治療剤組成物、抗炎症治療剤組成物、免疫治療剤組成物、癌の予防又は治療用組成物のいずれか1つであることを特徴とする、治療剤組成物。 - 前記治療剤組成物は、前記類似間葉系幹細胞から分泌される有効成分又はエクソソームをさらに含む、請求項10に記載の治療剤組成物。
- 前記治療剤組成物は、多発性硬化症、全身性硬化症、急性心筋梗塞、慢性心筋梗塞、慢性肺疾患、急性肺疾患、クローン病、便失禁、移植片対宿主病、下肢虚血疾患、閉塞性血栓性血管炎(Buerger病)、足部潰瘍、ループス、リウマチ性関節炎、急性及び慢性腎盂炎、炎症性膀胱炎、間質性膀胱炎、低活動性膀胱、過敏性膀胱、五十肩、回旋筋腱板損傷及び破裂、様々な運動などにより発生する筋骨格系損傷、膝軟骨損傷、耳鳴症、アトピー性皮膚炎、乾癬、火傷による皮膚損傷、紫外線などによる皮膚損傷、網膜症を含む炎症性及び免疫系疾患の予防、虚血性認知症、アルツハイマー認知症、脊髄損傷、パーキンソン病及び中枢神経系疾患のいずれか1つ以上の疾病の予防又は治療のためのものである、請求項10又は請求項11に記載の治療剤組成物。
- 請求項3~請求項9のいずれか一項に記載の類似間葉系幹細胞を含む伝達体であって、
前記伝達体は、医薬組成物を担持することを特徴とする、伝達体。 - 請求項3~請求項9のいずれか一項に記載の類似間葉系幹細胞から分泌される有効成分又はエクソソームを含む、疾病の予防又は治療用組成物。
- 前記疾病は、多発性硬化症、全身性硬化症、急性心筋梗塞、慢性心筋梗塞、慢性肺疾患、急性肺疾患、クローン病、便失禁、移植片対宿主病、下肢虚血疾患、閉塞性血栓性血管炎、足部潰瘍、ループス、リウマチ性関節炎、急性及び慢性腎盂炎、炎症性膀胱炎、間質性膀胱炎、低活動性膀胱、過敏性膀胱、五十肩、回旋筋腱板損傷及び破裂、様々な運動などにより発生する筋骨格系損傷、膝軟骨損傷、耳鳴症、アトピー性皮膚炎、乾癬、火傷による皮膚損傷、紫外線などによる皮膚損傷、網膜症を含む炎症性及び免疫系疾患の予防、虚血性認知症、アルツハイマー認知症、脊髄損傷、パーキンソン病及び中枢神経系疾患のいずれか1つ以上である、請求項14に記載の疾病の予防又は治療用組成物。
- 請求項3~請求項9のいずれか一項に記載の類似間葉系幹細胞から分泌される有効成分又はエクソソームを含む、化粧料組成物。
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