Osada Hiroaki, Kawatou Masahide, Fujita Daiki, Tabata Yasuhiko, Minatoya Kenji, Yamashita Jun K, Masumoto Hidetoshi
Department of Cardiovascular Surgery, Graduate School of Medicine, Kyoto University, Kyoto, Japan.
Department of Cell Growth and Differentiation, Center for iPS Cell Research and Application, Kyoto University, Kyoto, Japan.
JTCVS Open. 2021 Oct 1;8:359-374. doi: 10.1016/j.xjon.2021.09.038. eCollection 2021 Dec.
To establish a protocol to prepare and transplant clinical-grade human induced pluripotent stem cell (hiPSC)-derived cardiac tissues (HiCTs) and to evaluate the therapeutic potential in an animal myocardial infarction (MI) model.
We simultaneously differentiated clinical-grade hiPSCs into cardiovascular cell lineages with or without the administration of canonical Wnt inhibitors, generated 5- layer cell sheets with insertion of gelatin hydrogel microspheres (GHMs) (HiCTs), and transplanted them onto an athymic rat MI model. Cardiac function was evaluated by echocardiography and cardiac magnetic resonance imaging and compared with that in animals with sham and transplantation of 5-layer cell sheets without GHMs. Graft survival, ventricular remodeling, and neovascularization were evaluated histopathologically.
The administration of Wnt inhibitors significantly promoted cardiomyocyte (CM) ( < .0001) and vascular endothelial cell (EC) ( = .006) induction, which resulted in cellular components of 52.0 ± 6.1% CMs and 9.9 ± 3.0% ECs. Functional analyses revealed the significantly lowest left ventricular end-diastolic volume and highest ejection fraction in the HiCT group. Histopathologic evaluation revealed that the HiCT group had a significantly larger median engrafted area (4 weeks, GHM(-) vs HiCT: 0.4 [range, 0.2-0.7] mm vs 2.2 [range, 1.8-3.1] mm; = .005; 12 weeks, 0 [range, 0-0.2] mm vs 1.9 [range, 0.1-3.2] mm; = .026), accompanied by the smallest scar area and highest vascular density at the MI border zone.
Transplantation of HiCTs generated from clinical-grade hiPSCs exhibited a prominent therapeutic potential in a rat MI model and may provide a promising therapeutic strategy in cardiac regenerative medicine.
建立制备和移植临床级人诱导多能干细胞（hiPSC）来源的心脏组织（HiCTs）的方案，并在动物心肌梗死（MI）模型中评估其治疗潜力。
我们在给予或不给予经典Wnt抑制剂的情况下，将临床级hiPSCs同时分化为心血管细胞谱系，生成插入明胶水凝胶微球（GHMs）的5层细胞片（HiCTs），并将其移植到无胸腺大鼠MI模型上。通过超声心动图和心脏磁共振成像评估心脏功能，并与假手术组和移植无GHMs的5层细胞片的动物进行比较。通过组织病理学评估移植物存活、心室重构和新生血管形成情况。
Wnt抑制剂的使用显著促进了心肌细胞（CM）（P <.0001）和血管内皮细胞（EC）（P =.006）的诱导，导致细胞成分中CM占52.0 ± 6.1%，EC占9.9 ± 3.0%。功能分析显示，HiCT组的左心室舒张末期容积显著最低，射血分数最高。组织病理学评估显示，HiCT组的中位植入面积显著更大（4周时，无GHM组与HiCT组：0.4 [范围，0.2 - 0.7] mm对2.2 [范围，1.8 - 3.1] mm；P =.005；12周时，0 [范围，0 - 0.2] mm对1.9 [范围，0.1 - 3.2] mm；P =.026），同时MI边界区的瘢痕面积最小，血管密度最高。
临床级hiPSCs生成的HiCTs移植在大鼠MI模型中显示出显著的治疗潜力，可能为心脏再生医学提供一种有前景的治疗策略。
