Department of Pediatrics, Hualien Tzu Chi Hospital, Buddhist Tzu Chi Foundation and Tzu Chi University, Hualien.
Department of Medical Laboratory Science and Biotechnology, Asia University, Taichung.
Cell Transplant. 2022 Jan-Dec;31:9636897221085901. doi: 10.1177/09636897221085901.
There is an increasing interest in generating retinal pigment epithelial (RPE) cells from stem cells for treating degenerative eye diseases. However, whether human umbilical cord mesenchymal stem cells (HUCMSCs) can differentiate into RPE-like cells in a co-culture system has not been fully understood. In this study, induction of HUCMSC differentiation into RPE-like cells was performed by co-culturing HUCMSCs and a human RPE-like cell line (ARPE19) in a transwell system and then analyzed for biomarkers using quantitative reverse transcription polymerase chain reaction (RT-PCR) and immunofluorescence staining technique. Moreover, the functional characterization of induced cells was carried out by examining their phagocytic and neurotrophic factor-secreting activities. Our results showed that mRNA expressions of RPE-specific markers-MITF, OTX2, RPE65, PEDF, PME17, and CRALBP-and protein markers-RPE65, CRALBP, and ZO-1-were significantly increased in HUCMSC-derived RPE-like cells. Functional characteristic studies showed that these induced cells were capable of engulfing photoreceptor outer segments and secreting brain-derived neurotrophic factor (BDNF) and glial-derived neurotrophic factor (GDNF), which are typical functions of RPE-like cells. Overall, the study findings indicate that the morphology and proliferation of HUCMSCs can be maintained in a serum-free medium, and differentiation into RPE-like cells can be induced by simply co-culturing HUCMSCs with ARPE19 cells. Thus, the study provides fundamental information regarding the clinical-scale generation of RPE-like cells from HUCMSCs.
人们对于利用干细胞生成视网膜色素上皮（RPE）细胞治疗退行性眼病的兴趣日益浓厚。然而，人脐带间充质干细胞（HUCMSCs）是否能在共培养体系中分化为 RPE 样细胞尚未被充分了解。在这项研究中，我们通过将 HUCMSC 和人 RPE 样细胞系（ARPE19）共培养于 Transwell 系统中，诱导 HUCMSC 分化为 RPE 样细胞，然后使用实时定量聚合酶链反应（RT-PCR）和免疫荧光染色技术分析生物标志物。此外，我们还通过检测诱导细胞的吞噬和神经营养因子分泌活性，对其功能特征进行了研究。研究结果表明，在 HUCMSC 来源的 RPE 样细胞中，RPE 特异性标志物 MITF、OTX2、RPE65、PEDF、PME17 和 CRALBP 的 mRNA 表达以及 RPE65、CRALBP 和 ZO-1 等蛋白标志物的表达显著增加。功能特征研究表明，这些诱导细胞能够吞噬光感受器外节，并分泌脑源性神经营养因子（BDNF）和胶质源性神经营养因子（GDNF），这是 RPE 样细胞的典型功能。总的来说，该研究结果表明，在无血清培养基中可以维持 HUCMSC 的形态和增殖，并且通过简单地将 HUCMSC 与 ARPE19 细胞共培养即可诱导其分化为 RPE 样细胞。因此，该研究为从 HUCMSC 大规模生成 RPE 样细胞提供了基础信息。
