JP6164782B2 - 細胞外マトリックス材料から生理活性ゲルを製造する方法 - Google Patents
細胞外マトリックス材料から生理活性ゲルを製造する方法 Download PDFInfo
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Description
下記の例証では、例えば分離された膀胱粘膜下組織、小腸粘膜下組織、真皮のうちの1つ又は複数などの、ただしこれらに限定されない任意の数のECM製品を使用できる。下記の例証では、膀胱から分離され、上皮基底膜を有するECMであるUBMが、代表的なECMとして使用される。しかし、本明細書で開示される発明はUBMに限定されず、任意の分離されたECMに適用可能である。
Claims (17)
- 細胞外マトリックス材料から生理活性ゲルを製造する方法であって、
a)哺乳動物組織に由来し、天然型の前記組織で見出される量で配置されている、細胞外マトリックス材料の生理活性成分を含む、細胞外マトリックス材料(ECM)を用意することであって、前記ECMが、小腸粘膜下組織(SIS)、膀胱粘膜下組織(UBS)、膀胱マトリックス(UBM)及び肝臓基底膜(LBM)からなる群から選択され、
b)a)の前記細胞外マトリックス材料を微粒子化することと、
c)b)の前記微粒子化された細胞外マトリックス材料を、アルカリ性溶液中で可溶化することであって、前記可溶化が、前記微粒子化された細胞外マトリックス材料をゲル状へ転換し、
d)酸により、工程c)で調製されたゲルを中和することと
を含む、方法。 - e)工程d)で調製された、前記中和されたゲルを凍結することと、
f)工程e)で調製された、前記凍結したゲルを凍結乾燥することと
を更に含む、請求項1に記載の方法。 - 前記凍結乾燥したゲルを水溶液中で戻すことを更に含む、請求項2に記載の方法。
- 工程d)の後、4℃で最大48時間の滞留時間を更に含む、請求項1に記載の方法。
- 工程c)の時間が、30分〜48時間、2時間〜24時間、2日間〜3日間、3日間〜4日間、2日間〜7日間、及び3日間〜7日間からなる群から選択される、請求項1に記載の方法。
- 前記微粒子化された細胞外マトリックス材料の粒子範囲サイズが、1μm〜約1000μmである、請求項1に記載の方法。
- 前記細胞外マトリックス材料が、モル濃度範囲約0.1M〜約1.0MのNaOH中に、微粒子化された細胞外マトリックス材料の濃度約0.5%〜11%w/vで可溶化される、請求項1に記載の方法。
- 前記ゲルが、塩酸中で中和される、請求項1に記載の方法。
- 前記塩酸が、0.1M〜1.0Mの濃度を含む、請求項8に記載の方法。
- 前記生理活性成分が、線維芽細胞増殖因子−2(FGF−2)、結合組織増殖因子(CTGF)及び血管内皮増殖因子(VEGF)からなる群から選択される、請求項1に記載の方法。
- 前記細胞外マトリックスが2時間を超えて可溶化される際、前記可溶化された細胞外マトリックス材料中の前記線維芽細胞増殖因子−2(FGF−2)の濃度が、前記細胞外マトリックス材料が1.0MのNaOH中に可溶化される場合、100mMのNaOH中に可溶化される場合よりも高い、請求項10に記載の方法。
- 前記細胞外マトリックス材料が2時間を超えて可溶化される際、前記可溶化された細胞外マトリックス材料中の前記血管内皮増殖因子(VEGF)の濃度が、前記細胞外マトリックス材料が1.0MのNaOH中に可溶化される場合、100mMのNaOH中に可溶化される場合よりも高い、請求項10に記載の方法。
- 前記微粒子化された細胞外マトリックス材料がUBMを含み、前記UBMが、100mMのNaOH中に、微粒子化されたUBMの濃度7%で、4℃で可溶化される、請求項1に記載の方法。
- 工程(b)の前記微粒子化された材料が、200〜700ミクロンの範囲にある、請求項1に記載の方法。
- 微粒子化された細胞外マトリックス材料(ECM)を前記中和されたゲルに加えることをさらに含む、請求項1に記載の方法。
- 前記膀胱マトリックス(UBM)の可溶化が、数分から数時間の範囲の時間にわたる、請求項13に記載の方法。
- 前記範囲が、30分から12時間、12時間から24時間、24時間から36時間、36時間から48時間、2日間から7日間、及び3日間から7日間からなる群から選択される、請求項16に記載の方法。
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