Background: The human amniotic membrane (hAM) has long been utilized in ophthalmology for its regenerative and immunomodulatory properties. While its application in retinal repair is increasingly explored, particularly for macular holes, retinal tears, and age-related macular degeneration (AMD), most approaches emphasize the structural or paracrine support of hAM, often overlooking its cellular components. Human amniotic epithelial cells (hAECs), which naturally form a monolayer adherent to a basement membrane, closely resemble the architecture of the retinal pigment epithelium (RPE). This raises the possibility of leveraging native hAECs to generate RPE-like cell sheets directly on hAM for therapeutic use.

Methods: Term placentas were obtained from uncomplicated cesarean sections, with informed consent from the mothers. From these tissues, we compared hAECs cultured on their native hAM scaffold with hAECs enzymatically isolated and cultured on plastic. Cell morphology was examined under maintenance conditions, and the expression of selected RPE gene and protein markers was analyzed over time using qRT-PCR and immunofluorescence, without the use of differentiation inducers or exogenous signaling molecules.

Results: hAECs maintained on their native hAM scaffold exhibited improved epithelial integrity, reduced cellular stress, and more stable intercellular organization compared to isolated hAECs cultured on plastic. Several RPE markers were detectable in freshly harvested hAM, and their expression was better preserved over time in native cultures. Expression levels of both gene and protein markers varied among donor samples, indicating significant biological variability in the starting material.

Conclusions: These findings support the potential of hAECs as an accessible, ethically uncontroversial source for generating RPE-like cells. Based on these preliminary observations, it is now necessary to design and define appropriate differentiation protocols and establish standardization criteria for the selection and processing of donor material.