Enhanced recovery of mechanical function in the canine heart by seeding an extracellular matrix patch with mesenchymal stem cells committed to a cardiac lineage
Enhanced recovery of mechanical function in the canine heart by seeding an extracellular matrix patch with mesenchymal stem cells committed to a cardiac lineage
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DOI：
10.1152/ajpheart.00219.2008
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发表时间：
2008-12-01
影响因子：
4.8
通讯作者：
Cohen, Ira S.
中科院分区：
文献类型：
--
作者：
Potapova, Irina A.;Doronin, Sergey V.;Cohen, Ira S.
Potapova IA，Doronin SV，Kelly DJ，罗森AB，Schuldt AJ，Lu Z，Kochupura PV，罗宾逊RB，罗森MR，Brink PR，Gaudette GR，Cohen IS.通过向细胞外基质补片中接种定向分化为心脏谱系的间充质干细胞来增强犬心脏机械功能的恢复。Am J Physiol Heart Circ Physiol 295：H2257-H2263，2008。首次发表于2008年10月3日; doi：10.1152/ajpheart.00219.2008。再生组织的需要是至关重要的，特别是对于受伤后缺乏再生能力的心脏。膀胱细胞外基质（ECM），当用于修复右心室缺损，成功地再生了一些机械功能。本研究的目的是确定ECM的再生效果是否可以通过用增强的人间充质干细胞（hMSCs）接种贴片来改善，以分化出心脏谱系。hMSC用于形成三维球体。在暴露于球体形成的细胞中测定心脏蛋白的表达，并与未操作的hMSCs进行比较。为了确定是否存在功能性钙通道，将细胞膜片钳。为了评估这些细胞再生机械功能的能力，将球状体接种在ECM上，然后植入犬心脏以修复全层右心室缺损。结果，许多从球状体扩散的细胞表达心脏特异性蛋白，包括肌节α-辅肌动蛋白、心脏素和心房利钠肽，以及细胞周期标志物细胞周期蛋白D1和增殖细胞核抗原。16%的细胞中存在与心室肌细胞幅度相似的钙电流。与未经处理的hMSC-种子支架相比，心源性细胞-种子支架增加了犬心脏的局部机械功能。此外，用荧光标记物预标记的细胞表现出肌细胞特异性辅肌动蛋白染色，肌节间距与正常肌细胞相似。总之，球状体衍生的细胞表达心脏特异性蛋白，并表现出类似于成人心室肌细胞的钙电流。当这些细胞植入犬心脏时，其中一些细胞出现条纹，与未操作的hMSCs相比，机械功能得到改善。需要进一步的研究来确定机械功能的增加是否是由于心源性细胞向肌细胞的分化或其他效应。
Potapova IA, Doronin SV, Kelly DJ, Rosen AB, Schuldt AJ, Lu Z, Kochupura PV, Robinson RB, Rosen MR, Brink PR, Gaudette GR, Cohen IS. Enhanced recovery of mechanical function in the canine heart by seeding an extracellular matrix patch with mesenchymal stem cells committed to a cardiac lineage. Am J Physiol Heart Circ Physiol 295: H2257-H2263, 2008. First published October 3, 2008; doi: 10.1152/ajpheart.00219.2008. The need to regenerate tissue is paramount, especially for the heart that lacks the ability to regenerate after injury. The urinary bladder extracellular matrix (ECM), when used to repair a right ventricular defect, successfully regenerated some mechanical function. The objective of the current study was to determine whether the regenerative effect of ECM could be improved by seeding the patch with human mesenchymal stem cells (hMSCs) enhanced to differentiate down a cardiac linage. hMSCs were used to form three-dimensional spheroids. The expression of cardiac proteins was determined in cells exposed to the spheroid formation and compared with nonmanipulated hMSCs. To determine whether functional calcium channels were present, the cells were patch clamped. To evaluate the ability of these cells to regenerate mechanical function, the spheroids were seeded on ECM and then implanted into the canine heart to repair a full-thickness right ventricular defect. As a result, many of the cells spreading from the spheroids expressed cardiac-specific proteins, including sarcomeric alpha-actinin, cardiotin, and atrial natriuretic peptide, as well as the cell cycle markers cyclin D1 and proliferating cell nuclear antigen. A calcium current similar in amplitude to that of ventricular myocytes was present in 16% of the cells. The cardiogenic cell-seeded scaffolds increased the regional mechanical function in the canine heart compared with the unmanipulated hMSC-seeded scaffolds. In addition, the cells prelabeled with fluorescent markers demonstrated myocyte-specific actinin staining with sarcomere spacing similar to that of normal myocytes. In conclusion, the spheroid-derived cells express cardiac-specific proteins and demonstrate a calcium current similar to adult ventricular myocytes. When these cells are implanted into the canine heart, some of these cells appear striated and mechanical function is improved compared with the unmanipulated hMSCs. Further investigation will be required to determine whether the increased mechanical function is due to a differentiation of the cardiogenic cells to myocytes or to other effects.