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Nutrient microenvironments reprogram RPE metabolism.

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    The nutrient environment significantly impacts retinal pigment epithelium (RPE) cell function and metabolism. Choosing the right culture media is crucial for reproducible research in age-related macular degeneration (AMD) modeling.

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    Area of Science:

    • Cell Biology
    • Metabolic Engineering
    • Ophthalmology

    Background:

    • Induced pluripotent stem cell-derived retinal pigment epithelium (iPSC RPE) is vital for studying age-related macular degeneration (AMD).
    • Inconsistent RPE culture media composition hinders reproducible research on RPE metabolism and phenotype.

    Purpose of the Study:

    • To systematically investigate how six different nutrient microenvironments affect RPE phenotype, function, and metabolism.
    • To compare these effects in both iPSC RPE and fetal RPE (fRPE) models.

    Main Methods:

    • Cultured iPSC RPE and fRPE in six distinct media: MEMα, DMEM-HG/F12, HPLM+FBS, HPLM+B27, and X-VIVO 10.
    • Assessed RPE markers, cell morphology, transepithelial resistance, and metabolic profiles (amino acids, lipids, nucleotides).

    Main Results:

    • B27 and X-VIVO 10 media enhanced RPE cell size, hexagonality, and barrier function.
    • Specific media induced distinct metabolic changes: HPLM+FBS led to lipid accumulation, X-VIVO 10 caused vacuole formation, and B27 supplementation boosted respiration.
    • Metabolite analysis revealed condition-dependent shifts in consumption/production for creatine, serine, taurine, riboflavin, and guanine.

    Conclusions:

    • The nutrient microenvironment is a critical determinant of RPE phenotype, function, and metabolism.
    • This study provides essential data for selecting appropriate media and interpreting results in RPE disease modeling, particularly for AMD.