Culturing of Retinal Pigment Epithelium Cells

Valtink, Monika; Engelmann, Katrin

doi:10.1159/000223844

Cited by 14 publications

(8 citation statements)

References 44 publications

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“…Two main cell types were revealed: fl at polygonal epithelial cells in the center of colonies and elongated or fl attened fi broblastlike weakly or non-pigmented cells at their periphery (Fig 1, d, e). Similar morphotypes of RPE cells in vitro were described by other authors [5,6,17,21,27]. Some researchers believe that fi broblast-like morphology appears due to loss of contacts [4] and vast space for growth [16].…”

Section: Characteristics Of Intact Rpesupporting

confidence: 68%

Phenotypic Plasticity of Retinal Pigment Epithelial Cells from Adult Human Eye In Vitro

et al. 2011

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Phenotypic plasticity of retinal pigment epithelial cells from adult human eye was studied by immunohistochemical methods under different culturing conditions. It was found that retinal pigment epithelium in adult human eye is a heterogeneous population of cells demonstrating different behavior in vitro. Some cells retain epithelial morphology for a long time in culture, while others are rapidly transformed into fibroblast-like cells and synthesize proteins typical of proneural, neural, glial, and photoreceptor cells. However, irrespective of initial morphological features differentiation of retinal pigment cells can be modulated by varying culturing conditions.

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Section: Characteristics Of Intact Rpesupporting

confidence: 68%

Phenotypic Plasticity of Retinal Pigment Epithelial Cells from Adult Human Eye In Vitro

et al. 2011

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“…The maintenance of a differentiated state is linked to cell cycle arrest and when stimulating post-mitotic ocular cells to proliferate, e.g. by exposing them to culture conditions with a medium containing FBS, the cells may lose some of their differentiated characteristics (Valtink and Engelmann, 2009). Transitory cell dedifferentiation initially during primary culture is typical of differentiated cells (MacDonald, 1994).…”

Section: Discussionmentioning

confidence: 99%

Biopsy harvesting site and distance from the explant affect conjunctival epithelial phenotype ex vivo

Fostad

Eidet

Shatos

et al. 2012

Experimental Eye Research

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The purpose of the study was to investigate if the number of goblet cells expanded ex vivo from a conjunctival explant is affected by the biopsy harvesting site on the conjunctiva and the distance from the explant. Conjunctival explants from six regions: superior and inferior bulbus, fornix, and tarsus of male Sprague–Dawley rats were grown in RPMI 1640 with 10% fetal bovine serum on coverslips for eight days. Histochemical and immunofluorescent staining of goblet (CK-7/UEA-1/1MUC5AC), stratified squamous, non-goblet (CK-4), proliferating (PCNA) and progenitor (ABCG2) cells were analyzed by epifluorescence and laser confocal microscopy. Outgrowth was measured with NIH ImageJ. For statistical analysis the Mann–Whitney test and Spearman’s rank–order correlation test were used. Cultures from superior and inferior fornix contained the most goblet cells as indicated by the presence of CK-7+, UEA-1+ and MUC5AC+ cells. Superior and inferior forniceal cultures displayed 60.8% ± 9.2% and 64.7% ± 6.7% CK-7+ cells, respectively, compared to the superior tarsal (26.6% ± 8.4%; P < 0.05), superior bulbar (31.0% ± 4.0%; P < 0.05), inferior bulbar (38.5% ± 9.3%; P < 0.05) and inferior tarsal cultures (27.7% ± 8.3%; P < 0.05). While 28.4% ± 6.3% of CK-7+ goblet cells co-labeled with PCNA, only 7.4% ± 1.6% of UEA-1 + goblet cells did (P < 0.01). CK-7+ goblet cells were located at a lower concentration close to the explant (39.8% ± 3.1%) compared to near the leading edge (58.2% ± 4.5%; P < 0.05). Both markers for goblet cell secretory product (UEA-1 and MUC5AC), however, displayed the opposite pattern with a higher percentage of positive cells close to the explant than near the leading edge (P < 0.05). The percentage of CK-4+ cells was higher near the explant compared to near the leading edge (P < 0.01). The percentage of CK-7+ goblet cells in the cultures did not correlate with the outgrowth size (rs = −0.086; P = 0.435). The percentage of UEA-1 + goblet cells correlated negatively with outgrowth size (rs = −0.347; P < 0.01), whereas the percentage of CK-4+ cells correlated positively with the outgrowth size (rs = 0.473; P < 0.05). We conclude that forniceal explants yield the highest number of goblet cells ex vivo and thereby seem to be optimal for goblet cell transplantation. We also suggest that CK-7+/UEA-1− cells represent highly proliferative immature goblet cells. These cells could be important during conjunctival migration as they are mostly located close to the leading edge and their density does not decrease with increasing outgrowth size.

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“…To date, there exist a number of primary cell culture systems (fetal and adult) (Blenkinsop et al, 2013; Gamm et al, 2008; Hu and Bok, 2001; Maminishkis et al, 2006; Sonoda et al, 2009; Valtink and Engelmann, 2009) and cell lines (e.g., RPE-J, D407, and ARPE-19) (Davis et al, 1995; Dunn et al, 1996; Nabi et al, 1993) that have been used to evaluate basic RPE function as well as the molecular mechanisms underlying RPE-associated diseases. While the RPE culture system is convenient because it can be easily mass-produced and experimentally manipulated, validation of these culture models for known properties of RPE function and physiology is essential.…”

Section: Human Rpe Cell Culture As a Model For Studying Rpe Differmentioning

confidence: 99%

Cultured primary human fetal retinal pigment epithelium (hfRPE) as a model for evaluating RPE metabolism

Adijanto

Philp

2014

Experimental Eye Research

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Mitochondrial dysfunction has been shown to contribute to age-related and proliferative retinal diseases. Over the past decade, the primary human fetal RPE (hfRPE) culture model has emerged as an effective tool for studying RPE function and mechanisms of retinal diseases. This model system has been rigorously characterized and shown to closely resemble native RPE cells at the genomic and protein level, and that they are capable of accomplishing the characteristic functions of a healthy native RPE (e.g., rod phagocytosis, ion and fluid transport, and retinoid cycle). In this review, we demonstrated that the metabolic activity of the RPE is an indicator of its health and state of differentiation, and present the hfRPE culture model as a valuable in vitro system for evaluating RPE metabolism in the context of RPE differentiation and retinal disease.

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Culturing of Retinal Pigment Epithelium Cells

Cited by 14 publications

References 44 publications

Phenotypic Plasticity of Retinal Pigment Epithelial Cells from Adult Human Eye In Vitro

Phenotypic Plasticity of Retinal Pigment Epithelial Cells from Adult Human Eye In Vitro

Biopsy harvesting site and distance from the explant affect conjunctival epithelial phenotype ex vivo

Cultured primary human fetal retinal pigment epithelium (hfRPE) as a model for evaluating RPE metabolism

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