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Stereological estimation of cardiomyocyte number and proliferation
Methods ( IF 4.8 ) Pub Date : 2020-06-01 , DOI: 10.1016/j.ymeth.2020.06.002
Vasco Sampaio-Pinto 1 , Elsa D Silva 2 , Tiago L Laundos 3 , Paula da Costa Martins 4 , Perpétua Pinto-do-Ó 3 , Diana S Nascimento 3
Affiliation  

Cardiovascular diseases remain the leading cause of death, largely due to the limited regenerative capacity of the adult mammalian heart. Yet, neonatal mammals were shown to regenerate the myocardium after injury by increasing the proliferation of pre-existing cardiomyocytes. Re-activation of cardiomyocyte proliferation in adulthood has been considered a promising strategy to improve cardiac response to injury. Notwithstanding, quantification of cardiomyocyte proliferation, which occurs at a very low rate, is hampered by inefficient or unreliable techniques. Herein, we propose an optimized protocol to unequivocally assess cardiomyocyte proliferation and/or cardiomyocyte number in the myocardium. Resorting to a stereological approach we estimate the number of cardiomyocytes using representative thick sections of left ventricle fragments. This protocol overcomes the need for spatial-temporal capture of cardiomyocyte proliferation events by focusing instead on the quantification of the outcome of this process. In addition, assessment of cardiomyocyte nucleation avoids overestimation of cardiomyocyte proliferation due to increased binucleation. By applying this protocol, we were able to previously show that apical resection triggers proliferation of pre-existing cardiomyocytes generating hearts with more cardiomyocytes. Likewise, the protocol will be useful for any study aiming at evaluating the impact of neomyogenic therapies.

中文翻译:

心肌细胞数量和增殖的立体估计

心血管疾病仍然是导致死亡的主要原因,主要是由于成年哺乳动物心脏的再生能力有限。然而,新生哺乳动物在受伤后显示出通过增加预先存在的心肌细胞的增殖来再生心肌。成年期心肌细胞增殖的重新激活已被认为是改善心脏对损伤的反应的有希望的策略。尽管如此,以非常低的速率发生的心肌细胞增殖的量化受到低效或不可靠技术的阻碍。在此,我们提出了一种优化方案,以明确评估心肌中的心肌细胞增殖和/或心肌细胞数量。诉诸体视学方法,我们使用左心室碎片的代表性厚切片来估计心肌细胞的数量。该协议通过关注该过程结果的量化,克服了对心肌细胞增殖事件时空捕获的需要。此外,心肌细胞核化的评估避免了由于双核化增加而对心肌细胞增殖的高估。通过应用该协议,我们以前能够证明心尖切除会触发预先存在的心肌细胞的增殖,从而产生具有更多心肌细胞的心脏。同样,该协议对任何旨在评估新生肌治疗影响的研究都有用。心肌细胞成核的评估避免了由于双核化增加而高估心肌细胞增殖。通过应用该协议,我们以前能够证明心尖切除会触发预先存在的心肌细胞的增殖,从而产生具有更多心肌细胞的心脏。同样,该协议对任何旨在评估新生肌治疗影响的研究都有用。心肌细胞成核的评估避免了由于双核化增加而高估心肌细胞增殖。通过应用该协议,我们以前能够证明心尖切除会触发预先存在的心肌细胞的增殖,从而产生具有更多心肌细胞的心脏。同样,该协议对任何旨在评估新生肌治疗影响的研究都有用。
更新日期:2020-06-01
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