Shusterman Vladimir, Nagpal Prashant, Thedens Daniel, Zhu Xiaodong, Matasic Daniel S, Yoon Jin-Young, Morgan Gina, Hoffman Stacy, London Barry
The University of Iowa, Iowa City, IA.
PinMed, Inc., Pittsburgh, PA.
Biomed Phys Eng Express. 2019 Jul;5(4). doi: 10.1088/2057-1976/ab1c1c. Epub 2019 May 16.
Integrating cardiac-tissue patches into the beating heart and evaluating the long-term effects of such integration on cardiac contractility are two challenges in an emerging field of regenerative medicine. This pilot study presents tools for the imaging of contracting multicellular cardiac tissue constructs (MTCs) and demonstrates the feasibility of tracking the early development of strand geometry and contractions in ultrathin strands and layers of cardiac tissue using CINE MRI.
Cultured, ultrathin (~50-100-micron) MTCs of rat neonatal cardiomyocytes were plated in rectangular cell chambers (4.5 × 2.0 cm) with and without ultrathin, carbon EP electrodes embedded in the floor of the cell chamber. Two-dimensional, steady-state free precession (SSFP) CINE MRI, cell microscopy, and tissue photography were performed on Days 5-9 of cell development. Potential confounders and MRI artifacts were evaluated using non-contracting cardiac tissues and cell-free chambers filled with the cell-culture medium.
Synchronized contractions formed by Day 7; individual contracting tissue strands became identifiable by Day 9. The global patterns and details of the strand geometry and movement patterns in the SSFP images were in excellent agreement with microscopic and photographic images. No synchronized movement was identifiable by either microscopy or CINE MRI in the non-contracting MTCs or the cell-free medium. The EP recordings revealed well-defined depolarization and repolarization waveforms; the imaging artifacts generated by the carbon electrodes were small.
This pilot study demonstrates the feasibility of imaging cardiac-strand patterns and contractile activity in ultrathin, two-dimensional cardiac tissue in commonly used clinical scanners.
将心脏组织补片整合到跳动的心脏中,并评估这种整合对心脏收缩性的长期影响,是再生医学这一新兴领域面临的两大挑战。这项初步研究提供了用于对收缩的多细胞心脏组织构建体(MTC)进行成像的工具,并证明了使用电影磁共振成像(CINE MRI)追踪心脏组织超薄条带和层中条带几何形状的早期发育及收缩情况的可行性。
将培养的大鼠新生心肌细胞超薄(约50 - 100微米)MTC接种到矩形细胞培养室(4.5×2.0厘米)中,培养室底部嵌入或未嵌入超薄碳电极。在细胞发育的第5 - 9天进行二维稳态自由进动(SSFP)CINE MRI、细胞显微镜检查和组织摄影。使用不收缩的心脏组织和充满细胞培养基的无细胞培养室评估潜在的混杂因素和MRI伪影。
到第7天形成了同步收缩;到第9天可识别出单个收缩的组织条带。SSFP图像中条带几何形状和运动模式的整体模式及细节与显微镜和摄影图像高度吻合。在不收缩的MTC或无细胞培养基中,显微镜检查或CINE MRI均未发现同步运动。电极记录显示出明确的去极化和复极化波形;碳电极产生的成像伪影较小。
这项初步研究证明了在常用临床扫描仪中对超薄二维心脏组织中的心脏条带模式和收缩活动进行成像的可行性。