Allen Kristen, Pachter Natalie, Bandl Abigail, Qamar Haleema, Ropars Alex, Hookway Tracy A
Department of Biomedical Engineering, Binghamton University, The State University of New York, Binghamton, New York 13902, USA.
J Tissue Eng Regen Med. 2025 Jun 6;2025:3748093. doi: 10.1155/term/3748093. eCollection 2025.
Induced pluripotent stem cell-derived cardiomyocytes (iPSC-CMs) are used to model cardiac development and disease. This requires a robust population of mature CMs and external stimuli to mimic the complex environment of the heart. In effort toward this maturation, previous groups have applied electrical stimulation (ES) to CMs with varying results depending on the stimulation duration, frequency, and pattern. As such, there is an uncertainty surrounding the timeline on which stimulated iPSC-CMs begin to show early signs of maturation in comparison with their nonstimulated counterparts. Here, we introduce a low-cost custom bioreactor capable of delivering tunable ES to standard 2D cell monolayers. We show that, after exposure to short-term ES, stimulated CMs express early signs of maturation compared to nonstimulated controls. Changes to contractility and protein expression indicate cellular rearrangement within cell monolayers and induction of partial maturation in response to ES. While early signs of maturation are present after 3-4 days of ES, additional cellular structures must develop to reach complete maturation. We also show that this bioreactor can electrically stimulate cardiac fibroblasts (cFBs) and may induce alignment of cFB. We have shown that our custom ES bioreactor can be easily integrated into standard in vitro cell culture platforms to induce measurable changes in both CMs and cFB, exhibiting its potential for promoting crucial CM maturation and cell alignment for cardiac tissue engineering applications.
诱导多能干细胞衍生的心肌细胞（iPSC-CMs）被用于模拟心脏发育和疾病。这需要大量成熟的心肌细胞以及外部刺激来模拟心脏的复杂环境。为了实现这种成熟，先前的研究小组已将电刺激（ES）应用于心肌细胞，但结果因刺激持续时间、频率和模式而异。因此，与未受刺激的心肌细胞相比，受刺激的iPSC-CMs开始表现出早期成熟迹象的时间线仍存在不确定性。在这里，我们介绍了一种低成本的定制生物反应器，它能够向标准的二维细胞单层提供可调谐的电刺激。我们表明，在暴露于短期电刺激后，与未受刺激的对照组相比，受刺激的心肌细胞表现出早期成熟迹象。收缩性和蛋白质表达的变化表明细胞单层内的细胞重排以及对电刺激的部分成熟诱导。虽然在电刺激3-4天后出现了早期成熟迹象，但还必须发育出额外的细胞结构才能达到完全成熟。我们还表明，这种生物反应器可以对心脏成纤维细胞（cFBs）进行电刺激，并可能诱导cFB的排列。我们已经表明，我们的定制电刺激生物反应器可以很容易地集成到标准的体外细胞培养平台中，以诱导心肌细胞和成纤维细胞发生可测量的变化，展示了其在促进关键的心肌细胞成熟和细胞排列以用于心脏组织工程应用方面的潜力。
