Description
Purpose: Adult retinal pigment epithelial-19 (ARPE-19) cell line is widely used to study RPE physiology. Despite its common use, these cells show poor RPE-like characteristics including limited pigmentation and epithelial morphology. In the present study, we aimed to investigate the impact of the culture media on the morphology, pigmentation, and functional properties of the ARPE-19 cells, assessed by Ca2+ imaging.
Methods: ARPE-19 cells were seeded at a density of 30 000 cells/cm2 on a Matrigel-coated 96 well plates. The cells were cultured for 6 weeks either in DMEM supplemented with 1 % FBS and 1% Pen/Strep or in the commercially available XVIVO-10 medium. For live-cell Ca2+ imaging experiments, indo-1 was used as the Ca2+ indicator dye. Cellular responses were examined following stimulation with 100 µM ATP.
Results: ARPE-19 cells cultured in XVIVO-10 developed an RPE-like cobblestone morphology with evident pigmentation, whereas cells cultured in DMEM lacked both pigmentation and cobblestone morphology. All cells, regardless of culture conditions, exhibited Ca2+ responses following ATP stimulation. However, cells cultured in XVIVO-10 showed overall faster response kinetics, characterised by a more rapid rise and decline. In addition, the duration of the responses was significantly shorter in XVIVO-10-cultured cells. Cells cultured in DMEM recovered to baseline or near baseline Ca2+ levels after ATP stimulation, whereas responses in XVIVO-10-cultured cells plateaued at approximately half of the peak response.
Conclusions: ARPE-19 cells are typically cultured in DMEM. Our results indicate that culturing ARPE-19 cells in XVIVO-10 leads to improved RPE-like characteristics, including morphology and pigmentation. Moreover, Ca2+ responses were faster in X-VIVO10-cultured cells, suggesting an enhanced capacity for Ca2+ release. However, recovery was impaired, as Ca2+ levels failing to return to pre-stimulation baseline.
Lay Abstract
Researchers often use ARPE-19 cells as a model to study the retinal pigment epithelium (RPE), a layer of cells that supports vision. However, these cells do not always behave like real RPE cells when grown in the laboratory. In this study, we examined whether changing the cell culture medium could improve how closely these cells resemble those found in the human eye.
We compared two different culture media and found that one of them (X-VIVO 10) helped the cells develop more natural features, such as a typical shape and visible pigmentation. We also studied how the cells handled calcium, an important signal that helps regulate cell function. Cells grown in X-VIVO 10 responded more quickly to stimulation, but they did not fully return to their normal state afterward. These findings suggest that the choice of culture conditions can strongly influence how well ARPE-19 cells mimic real retinal cells. Improving these laboratory models may help researchers better study eye diseases and develop new treatments.
| Lay Title | How cell culture conditions affect the behavior of retinal cells in the laboratory |
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| Role | Postdoctoral Researcher |