Department of Science of Technology Innovation, Nagaoka University of Technology, 1603-1 Kamitomioka, Nagaoka, Niigata, 940-2188, Japan.
Department of Materials Science and Bioengineering, Nagaoka University of Technology, 1603-1 Kamitomioka, Nagaoka, Niigata, 940-2188, Japan.
Biochem Biophys Res Commun. 2023 Sep 24;674:190-198. doi: 10.1016/j.bbrc.2023.06.040. Epub 2023 Jun 14.
Cardiomyocytes derived from human induced pluripotent stem cells (hiPSCs) are a promising cell source for regenerative medicine and drug discovery. However, the use of animal models for studying human cardiomyocytes derived from hiPSCs in vivo is limited and challenging. Given the shared properties between humans and zebrafish, their ethical advantages over mammalian models, and their immature immune system that is rejection-free against xenografted human cells, zebrafish provide a suitable alternative model for xenograft studies. We microinjected fluorescence-labeled cardiac lineage cells derived from hiPSCs, specifically mesoderm or cardiac mesoderm cells, into the yolk and the area proximal to the outflow tract of the linear heart at 24 hours post-fertilization (hpf). The cells injected into the yolk survived and did not migrate to other tissues. In contrast, the cells injected contiguous with the outflow tract of the linear heart migrated into the pericardial cavity and heart. After 1 day post injection (1 dpi, 22-24 hpi), the injected cells migrated into the pericardial cavity and heart. Importantly, we observed heartbeat-like movements of some injected cells in the zebrafish heart after 1 dpi. These results suggested successful xenografting of hiPSC-derived cardiac lineage cells into the zebrafish embryo heart. Thus, we developed a valuable tool using zebrafish embryos as a model organism for investigating the molecular and cellular mechanisms involved in the grafting process. This is essential in developing cell transplantation-based cardiac therapeutics as well as for drug testing, notably contributing to advancements in the field of cardio-medicine.
人诱导多能干细胞（hiPSCs）衍生的心肌细胞是再生医学和药物发现的有前途的细胞来源。然而，在体内研究源自 hiPSC 的人类心肌细胞的动物模型的使用受到限制和具有挑战性。鉴于人类和斑马鱼之间的共同特性、它们相对于哺乳动物模型的伦理优势以及它们不成熟的免疫系统对异种移植的人类细胞无排斥反应，斑马鱼为异种移植研究提供了合适的替代模型。我们将荧光标记的源自 hiPSC 的心脏谱系细胞（特别是中胚层或心脏中胚层细胞）微注射到受精后 24 小时（hpf）的蛋黄和靠近线性心脏流出道的区域。注射到蛋黄中的细胞存活下来，没有迁移到其他组织。相比之下，注射到线性心脏流出道附近的细胞迁移到心包腔和心脏。在注射后 1 天（1 dpi，22-24 hpi），注射的细胞迁移到心包腔和心脏。重要的是，我们在 1 dpi 后观察到一些注射细胞在斑马鱼心脏中出现类似心跳的运动。这些结果表明 hiPSC 衍生的心脏谱系细胞成功异种移植到斑马鱼胚胎心脏中。因此，我们使用斑马鱼胚胎作为模型生物开发了一种有价值的工具，用于研究移植过程中涉及的分子和细胞机制。这对于开发基于细胞移植的心脏治疗方法以及药物测试至关重要，特别是为心脏医学领域的进步做出了贡献。
