Advantages of nanofibrous membranes for culturing of primary RPE cells compared to commercial scaffolds
The result's identifiers
Result code in IS VaVaI
<a href="https://www.isvavai.cz/riv?ss=detail&h=RIV%2F67985904%3A_____%2F22%3A00560631" target="_blank" >RIV/67985904:_____/22:00560631 - isvavai.cz</a>
Alternative codes found
RIV/61389013:_____/22:00560631 RIV/68378041:_____/22:00560631 RIV/00064173:_____/22:43922457 RIV/00216208:11120/22:43922457 and 2 more
Result on the web
<a href="https://asep.lib.cas.cz/arl-cav/cs/csg/?repo=crepo1&key=70763693065" target="_blank" >https://asep.lib.cas.cz/arl-cav/cs/csg/?repo=crepo1&key=70763693065</a>
DOI - Digital Object Identifier
<a href="http://dx.doi.org/10.1111/aos.15034" target="_blank" >10.1111/aos.15034</a>
Alternative languages
Result language
angličtina
Original language name
Advantages of nanofibrous membranes for culturing of primary RPE cells compared to commercial scaffolds
Original language description
Purpose Dysfunction of the retinal pigment epithelium (RPE) causes numerous forms of retinal degeneration. RPE replacement is a modern option to save vision. We aimed to test the results of transplanting cultured RPEs on biocompatible membranes. Methods We cultivated porcine primary RPE cells isolated from cadaver eyes from the slaughterhouse on two types of membranes: commercial polyester scaffolds Transwell (Corning Inc., Kenneburg, ME, USA) with 0.4 mu m pore size and prepared Poly (L-lactide-co-DL-lactide) (PDLLA) nanofibrous membranes with an average pore size of 0.4 mu m. Results Five types of assays were used for the analysis: immunocytochemistry (ICC), phagocytosis assay, Western blotting, real-time qPCR (RT-qPCR) and electron microscopy. RT-qPCR demonstrated that RPEs cultured on nanofibrous membranes have higher expressions of BEST1 (bestrophin 1), RLBP1 (retinaldehyde-binding protein 1), RPE65 (retinal pigment epithelium-specific 65 kDa protein), PAX6 (transcription factor PAX6), SOX9 (transcription factor SOX9), DCT (dopachrome tautomerase) and MITF (microphthalmia-associated transcription factor). ICC of the RPEs cultured on nanofibrous membranes showed more intensive staining of markers such as BEST1, MCT1 (monocarboxylate transporter 1), Na+/K(+)ATPase, RPE65 and acetylated tubulin in comparison with commercial ones. Additionally, the absence of alpha-SMA proved the stability of the RPE polarization state and the absence of epithelial-to-mesenchymal transition. RPE possessed high phagocytic activity. Electron microscopy of both membranes confirmed a confluent layer of RPE cells and their genuine morphological structure, which was comparable to native RPEs. Conclusions Retinal pigment epitheliums cultured on polylactide nanofibrous membranes improved the final quality of the cell product by having better maturation and long-term survival of the RPE monolayer compared to those cultured on commercial polyester scaffolds. PDLLA-cultured RPEs are a plausible source for the replacement of non-functioning RPEs during cell therapy.
Czech name
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Czech description
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Classification
Type
J<sub>imp</sub> - Article in a specialist periodical, which is included in the Web of Science database
CEP classification
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OECD FORD branch
30401 - Health-related biotechnology
Result continuities
Project
Result was created during the realization of more than one project. More information in the Projects tab.
Continuities
P - Projekt vyzkumu a vyvoje financovany z verejnych zdroju (s odkazem do CEP)
Others
Publication year
2022
Confidentiality
S - Úplné a pravdivé údaje o projektu nepodléhají ochraně podle zvláštních právních předpisů
Data specific for result type
Name of the periodical
Acta Ophthalmologica
ISSN
1755-375X
e-ISSN
1755-3768
Volume of the periodical
100
Issue of the periodical within the volume
5
Country of publishing house
DK - DENMARK
Number of pages
14
Pages from-to
"E1172"-"E1185"
UT code for WoS article
000710006200001
EID of the result in the Scopus database
2-s2.0-85117446785