Fight Against Angiogenesis-Related Blindness (FARB) Laboratory, Clinical Research Institute, Seoul National University Hospital, Seoul 03082, Republic of Korea.
Graduate School of Translational Medicine, Seoul National University College of Medicine, Seoul 03080, Republic of Korea.
Cells. 2023 Jun 16;12(12):1644. doi: 10.3390/cells12121644.
Marmosets have emerged as a valuable primate model in ophthalmic research due to their similarity to the human visual system and their potential for generating transgenic models to advance the development of therapies. In this study, we isolated and cultured primary retinal pigment epithelium (RPE) cells from marmosets to investigate the mechanisms underlying RPE dysfunction in aging and age-related macular degeneration (AMD). We confirmed that our culture conditions and materials supported the formation of RPE monolayers with functional tight junctions that closely resembled the in vivo RPE. Since serum has been shown to induce epithelial-mesenchymal transition (EMT) in RPE cells, we compared the effects of fetal bovine serum (FBS) with serum-free supplements B27 on transepithelial electrical resistance (TER), cell proliferation, and morphological characteristics. Additionally, we assessed the age-related morphological changes of in vivo and primary RPE cells. Our results indicate that primary marmoset RPE cells exhibit in vivo-like characteristics, while cells obtained from an older donor show evidence of aging, including a failure to form a polarized monolayer, low TER, and delayed cell cycle. In conclusion, our primary marmoset RPE cells provide a reliable in vitro model for developing novel therapeutics for visual-threatening disorders such as AMD, which can be used before animal experiments using marmosets.
食蟹猴在眼科研究中已经成为一种有价值的灵长类模型,因为它们的视觉系统与人类相似,并且有可能产生转基因模型来推进治疗方法的发展。在这项研究中,我们从食蟹猴中分离和培养了原代视网膜色素上皮 (RPE) 细胞,以研究衰老和年龄相关性黄斑变性 (AMD) 中 RPE 功能障碍的机制。我们证实,我们的培养条件和材料支持形成具有功能紧密连接的 RPE 单层,这些单层与体内 RPE 非常相似。由于血清已被证明会诱导 RPE 细胞发生上皮-间充质转化 (EMT),我们比较了胎牛血清 (FBS) 和无血清补充剂 B27 对跨上皮电阻 (TER)、细胞增殖和形态特征的影响。此外,我们评估了体内和原代 RPE 细胞的年龄相关形态变化。我们的结果表明,原代食蟹猴 RPE 细胞表现出与体内相似的特征,而来自较老供体的细胞则显示出衰老的迹象,包括无法形成极化单层、低 TER 和细胞周期延迟。总之,我们的原代食蟹猴 RPE 细胞为开发新型治疗 AMD 等视觉威胁性疾病的治疗方法提供了可靠的体外模型,可在使用食蟹猴进行动物实验之前使用。