Comparison of human amniotic membrane decellularisation approaches for hESC-derived RPE cells culture.
Comparison of human amniotic membrane decellularisation approaches for hESC-derived RPE cells culture.
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DOI：
10.1136/bmjophth-2022-000981
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发表时间：
2022-09
影响因子：
2.4
通讯作者：
Ponzin, Diego
中科院分区：
文献类型：
--
作者：
Daniele, Elena;Ferrari, Barbara;Rassu, Nicolo;Ben-Nun, Joshua;Bosio, Lorenzo;Barbaro, Vanessa;Ferrari, Stefano;Ponzin, Diego
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. Three enzymatic hAM de-epithelialisation methods (thermolysin, trypsin-EDTA and dispase II) were assessed by histological analysis and optical coherence tomography (OCT). We generated RPE cells from a human embryonic stem cell (hESC) line subjected to spontaneous differentiation in feeder-free conditions. The hESC-derived RPE cells were seeded over denuded hAM at a density of 2.0×105 cells/cm2 and maintained in culture for up to 4 weeks. Immnofluorescence was carried out to evaluate the development of a confluent monolayer of RPE cells on the top of the hAM. Conditioned medium was collected to measure pigment epithelium-derived factor (PEDF) concentration by ELISA. 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. 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.
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影响因子：
14
作者：
Ben M'Barek, Karim;Bertin, Stephane;Monville, Christelle
通讯作者：
Monville, Christelle
影响因子：
7.5
作者：
Hongisto H;Ilmarinen T;Vattulainen M;Mikhailova A;Skottman H
通讯作者：
Skottman H
影响因子：
4.3
作者：
Kiilgaard JF;Scherfig E;Prause JU;la Cour M
通讯作者：
la Cour M
DOI：
10.1089/clo.2006.8.189
发表时间：
2006-09-01
期刊：
CLONING AND STEM CELLS
影响因子：
--
作者：
Lund, Raymond D.;Wang, Shaomei;Lanza, Robert
通讯作者：
Lanza, Robert
影响因子：
4.4
作者：
Hopkinson, Andrew;McIntosh, Richard S.;Dua, Harminder S.
通讯作者：
Dua, Harminder S.