Comparison of human amniotic membrane decellularisation approaches for hESC-derived RPE cells culture

人类羊膜脱细胞方法对 hESC 衍生 RPE 细胞培养的比较

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作者:Elena Daniele #, Barbara Ferrari #, Nicolò Rassu, Joshua Ben-Nun, Lorenzo Bosio, Vanessa Barbaro, Stefano Ferrari, Diego Ponzin

Conclusion

The biological variability of the hAM could influence the adhesion and the expansion of hESC-derived RPE cells. Further studies are required to verify whether a non-confluent monolayer might represent a limit to transplantation.

Objective

Recent clinical studies have shown that the transplantation of functional retinal pigment epithelium (RPE) cells can prevent the onset of RPE degeneration in age-related macular degeneration. This study aimed to investigate the potential of human amniotic membrane (hAM) as a viable scaffold for the growth and proliferation of pluripotent-derived RPE cells.

Results

Laminin α5 and collagen IV staining confirmed the efficiency of the de-epithelialisation process. In particular, thermolysin showed good retention of tissue integrity on OCT images and greater preservation of the hAM basement membrane. The hESC-derived RPE cells formed patches of pigmented cells interspersed along the denuded hAM, but failed to form a regular sheet of RPE cells. These cells expressed typical RPE markers, such as PMEL17 and RPE65, but they secreted low levels of PEDF.

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