Purchase this article with an account.
Yini Wang, Jianing Gu, Qizhi Zhou, Jing Rao, Shibo Tang, Jiansu Chen; The Construction of Bioengineered RPE Sheet with Young and Macular RPE Characteristics. Invest. Ophthalmol. Vis. Sci. 2020;61(7):4134.
Download citation file:
© ARVO (1962-2015); The Authors (2016-present)
Retinal pigment epithelium (RPE) cells in macular display smaller, more regular hexagonal morphology and more melanosomes than those in peripheral retina. Developing a suitable tissue engineering (TE) sheet with young and macular RPE characteristics is beneficial for the cell therapy of age-related macular degeneration (AMD).
Primary RPE cells were isolated from human eyes balls. Adult RPE cells were cultured with normal medium (NM) and induced pluripotent stem cells conditioned medium (iPS-CM). And iPS cells derived from peripheral blood-derived cells were differentiated into RPE cells. Then iPS-RPE cells were seeded on femtosecond laser lntrastromal lenticule (FLI-lenticule) or tissue culture plate (TCP) with iPS-CM. Cultured RPE cells were assayed by immunofluorescence (IF), qPCR and RNA-seq.
iPS-CM significantly upregulated RPE progenitor-related genes, such as PMEL, TYRP1 and DCT genes, in cultured adult RPE cells (Fig. 1A). Adult RPE cells in iPS-CM could be confluent monolayers and polygonal shape with more compact and obvious hexagonal features than those in NM by ZO-1 staining (Fig. 1B). iPS cells enabled to be differentiate into RPE cells in induction medium. iPS-RPE cells presented plenty of melanin ( Fig. 2A). Adult RPE cells and iPS-RPE cells both displayed a typical polygonal morphology and expressed RPE specific phenotypes of RPE65, EMMPRIN and MITF in iPS-CM. Interestingly, iPS-RPE cells on FLI-lenticule revealed a distinctive, smaller and tighter epithelial phenotype compared to cells grown on TCP when cells of both groups were cultured with iPS-CM (Fig. 2B). At the same time, iPS-RPE cells significantly upregulated RPE progenitor related genes PMEL, TYRP1 and DCT on FLI-lenticule by qPCR (Fig. 2C).
This study demonstrates that iPS-CM in conjunction with FLI-lenticule is an promising way to construct an TE RPE sheet using adult RPE cells or iPS-RPE cells with young and macular RPE characteristics, which will be helpful for future transplantation to treat AMD.
This is a 2020 ARVO Annual Meeting abstract.
Fig.1 (A) The heatmaps of the different expression of RPE- and stem cell-related genes in iPS-CM vs NM. (B) The ZO-1 immunofluorescence images of adult RPE cells cultured with iPS-CM and NM.
Fig.2 (A) iPSC differentiation to RPE cells. (B) IF staining of ZO-1 and average cell area (p<0.001). (C) qPCR analysis of RPE progenitor related markers.
This PDF is available to Subscribers Only