Abstract
Purpose : The retinal pigment epithelium (RPE) is a crucial cell layer that supports the retina's health. Its deterioration can lead to severe vision problems, such as age-related macular degeneration (AMD) and inherited eye diseases. Stem cell-derived RPE cell transplantation is one of the most promising therapeutic approaches, requiring efficient and scalable production methods with preserved functionality of RPE cells.
Methods : This study used microfluidic emulsification to fabricate biodegradable poly(ε-caprolactone) (PCL) microcarriers. These carriers were engineered to enhance the expansion and differentiation of RPE cells derived from induced pluripotent stem cells (iPSCs). These microcarriers feature a smooth surface and porous internal structure, which aids cell attachment, growth, proper orientation, and maintaining RPE characteristics.
Results : Our results demonstrate that RPE cells cultured on these microcarriers maintain key morphological and functional characteristics comparable to those grown using traditional two-dimensional (2D) culture methods. The microcarriers mimic a natural environment, enhancing cellular activities such as pigmentation, expression of RPE-specific markers (e.g., RPE65, Ezrin, and ZO-1), and phagocytic function, which are crucial for the cells' therapeutic efficacy. In addition, their porous design allows for better nutrient and oxygen flow while reducing physical stress on cells, a common issue in standard 2D and dense microcarrier systems.
A key achievement of this study is the cryopreservation of RPE cells directly on PCL microcarriers, ensuring high viability and functional integrity upon thawing. This allows for effective long-term storage and transport, maintaining cell quality. Post-thaw tests confirm the cells' morphology, polarization, and metabolic activity, including effective photoreceptor outer segment phagocytosis, which is crucial for retinal health.
Conclusions : These PCL microcarriers offer a scalable and effective way to produce RPE cells with the potential for treating retinal diseases. This technology meets the need for “ready-to-use” RPE cells in regenerative medicine, especially in areas with high rates of AMD and similar conditions. Future efforts will focus on adapting this method for clinical use, aiming to improve and expand its applications in eye tissue engineering.
Methods : This study used microfluidic emulsification to fabricate biodegradable poly(ε-caprolactone) (PCL) microcarriers. These carriers were engineered to enhance the expansion and differentiation of RPE cells derived from induced pluripotent stem cells (iPSCs). These microcarriers feature a smooth surface and porous internal structure, which aids cell attachment, growth, proper orientation, and maintaining RPE characteristics.
Results : Our results demonstrate that RPE cells cultured on these microcarriers maintain key morphological and functional characteristics comparable to those grown using traditional two-dimensional (2D) culture methods. The microcarriers mimic a natural environment, enhancing cellular activities such as pigmentation, expression of RPE-specific markers (e.g., RPE65, Ezrin, and ZO-1), and phagocytic function, which are crucial for the cells' therapeutic efficacy. In addition, their porous design allows for better nutrient and oxygen flow while reducing physical stress on cells, a common issue in standard 2D and dense microcarrier systems.
A key achievement of this study is the cryopreservation of RPE cells directly on PCL microcarriers, ensuring high viability and functional integrity upon thawing. This allows for effective long-term storage and transport, maintaining cell quality. Post-thaw tests confirm the cells' morphology, polarization, and metabolic activity, including effective photoreceptor outer segment phagocytosis, which is crucial for retinal health.
Conclusions : These PCL microcarriers offer a scalable and effective way to produce RPE cells with the potential for treating retinal diseases. This technology meets the need for “ready-to-use” RPE cells in regenerative medicine, especially in areas with high rates of AMD and similar conditions. Future efforts will focus on adapting this method for clinical use, aiming to improve and expand its applications in eye tissue engineering.
| Original language | English |
|---|---|
| Journal | Investigative Ophthalmology and Visual Science |
| Volume | 66 |
| Issue number | 8 |
| Publication status | Published - 30 Jun 2025 |
