Studying the Morphology and Growth of Human Retinal Pigment Epithelial Cells on a Thin Layer of Alginate and Alginate/Gelatin as a Culture Substrate

Objective: Human retinal pigment epithelium (hRPE) is a cell monolayer located in the outer part of the retina that is in contact with photoreceptors. In many diseases RPE cells damage. One way for treating this disease is the implantation of intact instead of damaged cells. For this reason different types of substrates have been used for cell cultivation. This study has used alginate and a blend of alginate/gelatin (A/G) to study RPE cell growth. Methods: We prepared alginate solutions in concentrations of ۱% and ۲% (w/v) in water and DMEM/F۱۲. The solutions were infused into each well of ۶-well micro plates until a uniform culture substrate that had a ۱ mm thickness was generated. Passage-۴ hRPE cells were cultivated on the substrate and the cell characteristics studied. hRPE cells did not adhere to alginate in DMEM/F۱۲ and did not exhibit interaction with alginate substrate. For this reason A/G solutions at concentrations of ۱% and ۲% (w/v) in water were prepared. We prepared A/G blends at weight ratios of: ۲۰:۸۰, ۳۰:۷۰, ۴۰:۶۰, ۵۰:۵۰, ۶۰:۴۰, ۷۰:۳۰, and ۸۰:۲۰. These blends were infused into each well of ۶-well plates until the appropriate ۱ mm thickness of A/G was achieved. Isolated hRPE cells were cultured on synthetic substrate after which we studied the cells' characteristics.   Result: hRPE cell generated adhesive colonies on the A/G substrate. In all studied combinations of A/G, the diffused hRPE cells formed a monolayer under the substrate sheets. However the A/G ۲۰:۸۰ ratio had cell growth in the upper face of the substrate. hRPE survived indefinitely on A/G substrate. After the cells were re-cultured on polystyrene, they showed general morphological features of normal hRPE cells. Conclusion: The A/G blend at a ۲۰:۸۰ ratio was chosen to be used for future studies.

کلیدواژه ها:

Tissue engineering ، Human retinal pigment epithelium cell ، Substrate ، Alginate and gelatin ، مهندسی بافت ، سلولهای رنگدانهدار شبکیه انسان ، بستر ، آلژینات ، ژلاتین