JP2021121206A - 筋細胞パッチおよびその使用 - Google Patents
筋細胞パッチおよびその使用 Download PDFInfo
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Abstract
Description
本願は、参照によりその全体が本明細書に援用される、2013年10月9日出願の米国仮特許出願第61/888882号の優先権を主張する。
本発明は、VAによって与えられた助成金番号1−101−BX001406−01A1の下で政府支援によってなされたものである。
(a)線維芽細胞含を含む3次元足場(3DFCS)の表面上に未成熟収縮細胞を播種して、収縮構築物を生成することと、
(b)条線を形成する成熟収縮細胞への未成熟収縮細胞の分化を促進する条件下で収縮構築物を培養することと、
を含む方法を提供する。
・形態学的に小さい細胞サイズ;
・筋原繊維密度の減少;
・電気生理学的に抑えられ/減少した活動電位振幅;
・MYH7(ベータミオシン重鎖)、MYH6(アルファミオシン重鎖)、SCN5A、GJA1(コネキシン43)、HCN4(過分極活性化K+チャネル)、KCNJ2(内向き整流性カリウムイオンチャネル)、SERCA2a(サルコ/小胞体(sarcoendoplasmic reticulum)ATPアーゼ)、アルファアクチニン、心臓トロポニンI(cTnI)、心臓トロポニンT(cTnT)の遺伝子および/またはタンパク質の発現の減少。
・形態学的に小さい細胞サイズ;
・筋原繊維密度の減少;
・電気生理学的に抑えられ/減少した活動電位振幅;
・MYH7(ベータミオシン重鎖)、MYH6(アルファミオシン重鎖)、SCN5A、GJA1(コネキシン43)、HCN4(過分極活性化K+チャネル)、KCNJ2(内向き整流性カリウムイオンチャネル)、SERCA2a(サルコ/小胞体ATPアーゼ)、アルファアクチニン、心臓トロポニンI(cTnI)、心臓トロポニンT(cTnT)の遺伝子および/またはタンパク質の発現の減少。
クラスI:いかなる活動でも制約を経験しない;通常の活動からの症状はない。
クラスII:活動にわずかに軽い制約がある;患者は安静時または軽度の労作時に快適である。
クラスIII:いかなる活動でも顕著な制約がある;患者は安静時にのみ快適である。
クラスIV:いかなる身体活動も不快感をもたらし、安静時に症状が起こる。
(a)収縮構築物を生成するための線維芽細胞含を含む3次元足場(3DFCS)の表面上に未成熟収縮細胞を播種することと、
(b)条線を形成する成熟収縮細胞への未成熟収縮細胞の分化を促進する条件下で、収縮構築物を培養することと、
を含む方法を提供する。
パッチの製造
播種方法
簡単に説明すると、懸濁液中の細胞に遠心力を適用する。3次元線維芽細胞構築物(3DFC)の表面上に細胞を押しやる/強いると、細胞をランダムだが均一に分布できる。ベースの構築物(3DFC)が細胞の移植および配置のための支持体ならびに適切な要件を提供すると、収縮力が生じる。最終「生成物」は、線維芽細胞が埋め込まれ、収縮細胞集団、この調製ではiPSC由来の(「未成熟」)心筋細胞が播種された分解性メッシュである。
誘導性ヒト多能性幹細胞(hiPSC)の播種密度は、0.3×106細胞/cm2〜2.4×106細胞/cm2の範囲であり、1.2×106細胞/cm2が理想である。
心臓パッチ用の出発材料は、ヒト皮膚線維芽細胞が埋め込まれた合成ビクリルメッシュを含む3DFCである。線維芽細胞は血管新生を示し、したがって、心臓への移植後、播種されたiPSC由来の心筋細胞集団に栄養サポートを提供する。発明者らのデータは、細胞比(皮膚線維芽細胞に対するiPSC由来の心筋細胞)が3:20〜1.2:1の範囲であり、1.2:2が理想であることを示している。
1. Thai, H. M., Juneman, E., Lancaster, J., Hagerty, T., Do, R., Castellano, L., Kellar, R., Williams, S., Sethi, G., Schmelz, M., Gaballa, M., & Goldman, S. (2009). Implantation of a three-dimensional fibroblast matrix improves left ventricular function and blood flow after acute myocardial infarction. Cell Transplant., 18(3), 283-295. PMC2739416:PM19558777. doi:10.3727/096368909788535004
2. Lancaster, J., Juneman, E., Hagerty, T., Do, R., Hicks, M., Meltzer, K., Standley, P., Gaballa, M., Kellar, R., Goldman, S., & Thai, H. (2010). Viable fibroblast matrix patch induces angiogenesis and increases myocardial blood flow in heart failure after myocardial infarction. Tissue Eng.Part A., 16(10), 3065-3073. PM20486785. doi:10.1089/ten.TEA.2009.0589
3. Lancaster, J. J., Arnce, S. A., Johnson, N. M., Juneman, E. B., Thai, H. M., Kellar, R. S., Vitorin, J. E., Burt, J. M., Bahl, J. J., & Goldman, S. (2010). In vivo evaluation of a biologically active cardiomyocyte seeded scaffold to treat chronic heart failure. [abstract]. Paper presented at the Heart Failure Society of America: 14th Annual Scientific Meeting, San Diego,CA. , 16(8) S45. doi:10.1016/j.cardfail.2010.06.155
4. Lancaster, J. J., Arnce, S. A., Johnson, N. M., Juneman, E. B., Thai, H. M., Kellar, R. S., Vitorin, J. E., Burt, J. M., Gaballa, M. A., Bahl, J. J., & Goldman, S. Tissue engineered scaffold seeded with cardiomyocytes improves cardiac function in rats with chronic ischemic heart failure disease. (Journal of Heart and Lung Transplantation).
Claims (1)
- 収縮構築物を調製するための方法であって、
(a)線維芽細胞含を含む3次元足場(3DFCS)の表面上に未成熟収縮細胞を播種して、収縮構築物を生成することと、
(b)条線を形成する成熟収縮細胞への未成熟収縮細胞の分化を促進する条件下で前記収縮構築物を培養することと、
を含む方法。
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