Enhanced gap junction expression in myoblast-containing engineered tissue.
Enhanced gap junction expression in myoblast-containing engineered tissue.
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DOI：
10.1016/j.bbrc.2012.05.016
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发表时间：
2012-06-08
影响因子：
3.1
通讯作者：
Choi YH
中科院分区：
文献类型：
--
作者：
Srinivasan SP;Neef K;Treskes P;Liakopoulos OJ;Stamm C;Cowan DB;Madershahian N;Kuhn E;Slottosch I;Wittwer T;Wahlers T;Choi YH
骨骼肌成肌细胞移植是一种潜在的心肌细胞治疗方法。SM是可自体获得的，预定用于肌肉分化和抗缺血。其临床应用的主要障碍是细胞分离期间纯度和产量的限制以及分化成肌管后缺乏间隙连接表达。此外，移植的SM不与宿主心肌功能或电整合。在这里，我们描述了一种有效的方法分离同质SM人口从新生小鼠，并表现出持久的间隙连接表达的工程组织。该方法在培养10天后从162.12 ± 11.85 mg肌肉组织中产生1.4 × 108个高纯度SM（>99%结蛋白阳性）。血清饥饿条件下，有效地诱导分化成自发收缩肌管，正好与间隙连接表达的损失。对于机械调节，将细胞整合到工程化组织构建体中。组织构建体内的SM表现出长期存活、有序排列和保留的分化成收缩性肌管的能力。当组织结构受到被动纵向拉伸应力时，间隙连接和细胞粘附蛋白的表达在整个分化过程中保持或增加。我们的研究表明，SM的机械负载可以提供心肌内改善的机电整合，这可能导致更多的治疗机会。
Transplantation of skeletal myoblasts (SMs) has been investigated as a potential cardiac cell therapy approach. SM are available autologously, predetermined for muscular differentiation and resistant to ischemia. Major hurdles for their clinical application are limitations in purity and yield during cell isolation as well as the absence of gap junction expression after differentiation into myotubes. Furthermore, transplanted SMs do not functionally or electrically integrate with the host myocardium. Here, we describe an efficient method for isolating homogeneous SM populations from neonatal mice and demonstrate persistent gap junction expression in an engineered tissue. This method resulted in a yield of 1.4 × 108 high-purity SMs (>99% desmin positive) after 10 days in culture from 162.12 ± 11.85 mg muscle tissue. Serum starvation conditions efficiently induced differentiation into spontaneously contracting myotubes that coincided with loss of gap junction expression. For mechanical conditioning, cells were integrated into engineered tissue constructs. SMs within tissue constructs exhibited long term survival, ordered alignment, and a preserved ability to differentiate into contractile myotubes. When the tissue constructs were subjected to passive longitudinal tensile stress, the expression of gap junction and cell adherence proteins was maintained or increased throughout differentiation. Our studies demonstrate that mechanical loading of SMs may provide for improved electromechanical integration within the myocardium, which could lead to more therapeutic opportunities.
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影响因子：
37.8
作者：
Hofmann, M;Wollert, KC;Drexler, H
通讯作者：
Drexler, H
DOI：
10.1073/pnas.1232447100
发表时间：
2003-06-24
影响因子：
11.1
作者：
Léobon, B;Garcin, I;Charpak, S
通讯作者：
Charpak, S
影响因子：
37.8
作者：
Menasche, Philippe;Alfieri, Ottavio;Hagege, Albert A.
通讯作者：
Hagege, Albert A.
影响因子：
64.8
作者：
Laflamme, Michael A.;Murry, Charles E.
通讯作者：
Murry, Charles E.
影响因子：
5.2
作者：
Blocklet, Didier;Toungouz, Michel;Goldman, Serge
通讯作者：
Goldman, Serge