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

Area of Science:

• Regenerative Medicine
• Ophthalmology
• Stem Cell Biology

Background:

• Clinical studies suggest retinal pigment epithelium (RPE) cell transplantation can prevent degeneration in age-related macular degeneration (AMD).
• Human amniotic membrane (hAM) is being explored as a scaffold for cell transplantation therapies.

Purpose of the Study:

• To assess human amniotic membrane (hAM) as a scaffold for pluripotent-derived RPE cells.
• To evaluate RPE cell growth, proliferation, and function on de-epithelialized hAM.

Main Methods:

• Enzymatic de-epithelialization of hAM using thermolysin, trypsin-EDTA, and dispase II.
• Generation of RPE cells from human embryonic stem cells (hESCs).
• Seeding hESC-derived RPE cells onto hAM and culturing for 4 weeks, followed by immunofluorescence and PEDF measurement.

Main Results:

• Thermolysin treatment preserved hAM integrity and basement membrane structure.
• hESC-derived RPE cells formed pigmented patches but not a confluent monolayer on hAM.
• RPE cells expressed key markers (PMEL17, RPE65) but secreted low levels of PEDF.

Conclusions:

• Biological variability of hAM may affect hESC-derived RPE cell adhesion and expansion.
• Further investigation is needed to determine if a non-confluent RPE monolayer is suitable for transplantation in AMD.