From the Department of Bioengineering, University of Colorado Denver/Anschutz Medical Campus, Aurora, Colorado.
Department of Biomedical Engineering, University of Houston, Houston, Texas.
ASAIO J. 2018 Sep/Oct;64(5):e105-e114. doi: 10.1097/MAT.0000000000000765.
Bioengineering cardiac tissue constructs with adult cardiomyocytes may help treat adult heart defects and injury. In this study, we fabricated cardiac tissue constructs by seeding adult rat cardiomyocytes on a fibrin gel matrix and analyzed the electromechanical properties of the formed cardiac tissue constructs. Adult rat cardiomyocytes were isolated with a collagenase type II buffer using an optimized Langendorff perfusion system. Cardiac tissue constructs were fabricated using either indirect plating with cardiomyocytes that were cultured for 1 week and dedifferentiated or with freshly isolated cardiomyocytes. The current protocol generated (3.1 ± 0.5) × 10 (n = 5 hearts) fresh cardiomyocytes from a single heart. Tissue constructs obtained by both types of plating contracted up to 30 days, and electrogram (ECG) signals and contractile twitch forces were detected. The constructs bioengineered by indirect plating of dedifferentiated cardiomyocytes produced an ECG R wave amplitude of 15.1 ± 5.2 µV (n = 7 constructs), a twitch force of 70-110 µN, and a spontaneous contraction rate of about 390 bpm. The constructs bioengineered by direct plating of fresh cardiomyocytes generated an ECG R wave amplitude of 6.3 ± 2.5 µV (n = 8 constructs), a twitch force of 40-60 µN, and a spontaneous contraction rate of about 230 bpm. This study successfully bioengineered three-dimensional cardiac tissue constructs using primary adult cardiomyocytes.
用成体心肌细胞构建生物工程心脏组织可能有助于治疗成人心脏缺陷和损伤。在这项研究中,我们通过将成年大鼠心肌细胞接种在纤维蛋白凝胶基质上来构建心脏组织构建体,并分析了形成的心脏组织构建体的机电特性。使用优化的 Langendorff 灌注系统,用胶原酶 II 缓冲液分离成年大鼠心肌细胞。通过间接接种培养 1 周并去分化的心肌细胞或新鲜分离的心肌细胞来构建心脏组织构建体。该方案从单个心脏中产生了(3.1 ± 0.5)×10(n = 5 个心脏)个新鲜心肌细胞。两种接种方式获得的组织构建体均可收缩长达 30 天,并可检测到心电图(ECG)信号和收缩抽搐力。通过去分化心肌细胞间接接种生物工程化的构建体产生的 ECG R 波幅度为 15.1 ± 5.2 µV(n = 7 个构建体),抽搐力为 70-110 µN,自发收缩率约为 390 bpm。通过直接接种新鲜心肌细胞生物工程化的构建体产生的 ECG R 波幅度为 6.3 ± 2.5 µV(n = 8 个构建体),抽搐力为 40-60 µN,自发收缩率约为 230 bpm。本研究成功地使用原代成体心肌细胞构建了三维心脏组织构建体。