Shao Zhihua, Wang Haiyun, Zhou Xuejian, Guo Baosen, Gao Xuehu, Xiao Zengrong, Liu Meng, Sha Jihong, Jiang Chunlian, Luo Yuping, Liu Zhixue, Li Siguang
Stem Cell Translational Research Center, Tongji Hospital, Tongji University School of Medicine, Shanghai, China.
Shanghai First Maternity and Infant Health Hospital, Tongji University School of Medicine, Shanghai, China.
Cell Prolif. 2017 Dec;50(6). doi: 10.1111/cpr.12386. Epub 2017 Sep 18.
Primary retinal pigment epithelium (RPE) cells have a limited capacity to re-establish epithelial morphology and to maintain native RPE function in vitro, and all commercially available RPE cell lines have drawbacks of morphology or function; therefore, the establishment of new RPE cell lines with typical characteristics of RPE would be helpful in further understanding of their physiological and pathological mechanisms. Here, we firstly report a new spontaneously generated RPE line, fhRPE-13A, from a 13-week aborted foetus. We aimed to investigate its availability as a RPE model.
RNA-seq data were mapped to the human genome assembly hg19. Global transcriptional data were analysed by Weighted Gene Co-expression Network Analysis (WGCNA) and differentially expressed genes (DEGs). The morphology and molecular characteristics were examined by immunofluorescence, transmission electron micrographs, PCR and western blot. Photoreceptor outer segments (POS) phagocytosis assay and transepithelial resistance measurement (TER) were performed to assess phagocytic activity and barrier function, respectively.
The fhRPE-13A cells showed typical polygonal morphology and normal biological processes of RPE. Meanwhile they were capable of POS phagocytosis in vitro, and the expression level of TYR and TYRP1 were significantly higher than that in ARPE-19 cells.
The foetal human RPE line fhRPE-13A is a valuable system for researching phagocytosis and morphogenesis of RPE in vitro.
原代视网膜色素上皮(RPE)细胞在体外重建上皮形态和维持天然RPE功能的能力有限,并且所有市售的RPE细胞系都存在形态或功能方面的缺陷;因此,建立具有RPE典型特征的新RPE细胞系将有助于进一步了解其生理和病理机制。在此,我们首次报道了一种从13周流产胎儿中自发产生的新RPE细胞系fhRPE - 13A。我们旨在研究其作为RPE模型的可用性。
RNA测序数据被映射到人类基因组组装hg19。通过加权基因共表达网络分析(WGCNA)和差异表达基因(DEG)对全局转录数据进行分析。通过免疫荧光、透射电子显微镜、PCR和蛋白质免疫印迹法检测细胞的形态和分子特征。分别进行光感受器外段(POS)吞噬试验和跨上皮电阻测量(TER)以评估吞噬活性和屏障功能。
fhRPE - 13A细胞呈现典型的多边形形态和RPE正常的生物学过程。同时,它们在体外能够进行POS吞噬,并且TYR和TYRP1的表达水平显著高于ARPE - 19细胞。
胎儿人RPE细胞系fhRPE - 13A是体外研究RPE吞噬作用和形态发生的有价值的系统。