Advantages of nanofibrous membranes for culturing of primary
RPE cells compared to commercial scaffolds

J 2022

Advantages of nanofibrous membranes for culturing of primary RPE cells compared to commercial scaffolds

TICHOTOVÁ, Lucie, Hana STUDENOVSKÁ, Goran PETROVSKI, Štěpán POPELKA, Yaroslav NEMESH et. al.

Základní údaje

Originální název

Advantages of nanofibrous membranes for culturing of primary RPE cells compared to commercial scaffolds

Autoři

TICHOTOVÁ, Lucie (203 Česká republika), Hana STUDENOVSKÁ (203 Česká republika), Goran PETROVSKI, Štěpán POPELKA (203 Česká republika), Yaroslav NEMESH, Miroslava SEDLÁČKOVÁ (203 Česká republika, domácí), Saskia DRUTOVIČ, Sonali ROHIWAL, Pavla JENDELOVÁ (203 Česká republika), Slaven ERCEG, Anna BRYMOVÁ (203 Česká republika), Ana ARTERO-CASTRO, Lyubomyr LYTVYNCHUK, Zbyněk STRAŇÁK (203 Česká republika), Zdeňka ELLEDEROVÁ, Jan MOTLÍK a Taras ARDAN (garant)

Vydání

Acta Ophthalmologica, Hoboken, Wiley, 2022, 1755-375X

Další údaje

Jazyk

angličtina

Typ výsledku

Článek v odborném periodiku

Obor

30207 Ophthalmology

Stát vydavatele

Spojené státy

Utajení

není předmětem státního či obchodního tajemství

Odkazy

URL

Impakt faktor

Impact factor: 3.400

Kód RIV

RIV/00216224:14110/22:00125041

Organizační jednotka

Lékařská fakulta

DOI

http://dx.doi.org/10.1111/aos.15034

UT WoS

000710006200001

Klíčová slova anglicky

eye; nanoﬁbrous membrane; retina; retinal pigment epithelium; RPE

Štítky

14110517, rivok

Příznaky

Mezinárodní význam, Recenzováno

Změněno: 20. 7. 2022 08:14, Mgr. Tereza Miškechová

Anotace

V originále

ABSTRACT 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 µm pore size and prepared Poly (L-lactide-co-DL-lactide) (PDLLA) nanofibrous membranes with an average pore size of 0.4 µm. 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 α-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.

Citovat

TICHOTOVÁ, Lucie, Hana STUDENOVSKÁ, Goran PETROVSKI, Štěpán POPELKA, Yaroslav NEMESH, Miroslava SEDLÁČKOVÁ, Saskia DRUTOVIČ, Sonali ROHIWAL, Pavla JENDELOVÁ, Slaven ERCEG, Anna BRYMOVÁ, Ana ARTERO-CASTRO, Lyubomyr LYTVYNCHUK, Zbyněk STRAŇÁK, Zdeňka ELLEDEROVÁ, Jan MOTLÍK a Taras ARDAN. Advantages of nanofibrous membranes for culturing of primary RPE cells compared to commercial scaffolds. Acta Ophthalmologica. Hoboken: Wiley, 2022, roč. 100, č. 5, s. "E1172"-"E1185", 14 s. ISSN 1755-375X. Dostupné z: https://dx.doi.org/10.1111/aos.15034.

@article{1802211,
   author = {Tichotová, Lucie and Studenovská, Hana and Petrovski, Goran and Popelka, Štěpán and Nemesh, Yaroslav and Sedláčková, Miroslava and Drutovič, Saskia and Rohiwal, Sonali and Jendelová, Pavla and Erceg, Slaven and Brymová, Anna and ArteroandCastro, Ana and Lytvynchuk, Lyubomyr and Straňák, Zbyněk and Ellederová, Zdeňka and Motlík, Jan and Ardan, Taras},
   article_location = {Hoboken},
   article_number = {5},
   doi = {http://dx.doi.org/10.1111/aos.15034},
   keywords = {eye; nanoﬁbrous membrane; retina; retinal pigment epithelium; RPE},
   language = {eng},
   issn = {1755-375X},
   journal = {Acta Ophthalmologica},
   title = {Advantages of nanofibrous membranes for culturing of primary RPE cells compared to commercial scaffolds},
   url = {https://onlinelibrary.wiley.com/doi/epdf/10.1111/aos.15034},
   volume = {100},
   year = {2022}
}

TY  - JOUR
ID  - 1802211
AU  - Tichotová, Lucie - Studenovská, Hana - Petrovski, Goran - Popelka, Štěpán - Nemesh, Yaroslav - Sedláčková, Miroslava - Drutovič, Saskia - Rohiwal, Sonali - Jendelová, Pavla - Erceg, Slaven - Brymová, Anna - Artero-Castro, Ana - Lytvynchuk, Lyubomyr - Straňák, Zbyněk - Ellederová, Zdeňka - Motlík, Jan - Ardan, Taras
PY  - 2022
TI  - Advantages of nanofibrous membranes for culturing of primary RPE cells compared to commercial scaffolds
JF  - Acta Ophthalmologica
VL  - 100
IS  - 5
SP  - "E1172"-"E1185"
EP  - "E1172"-"E1185"
PB  - Wiley
SN  - 1755375X
KW  - eye
KW  - nanoﬁbrous membrane
KW  - retina
KW  - retinal pigment epithelium
KW  - RPE
UR  - https://onlinelibrary.wiley.com/doi/epdf/10.1111/aos.15034
N2  - ABSTRACT 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 µm pore size and prepared Poly (L-lactide-co-DL-lactide) (PDLLA) nanofibrous membranes with an average pore size of 0.4 µm. 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 α-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.
ER  -

TICHOTOVÁ, Lucie, Hana STUDENOVSKÁ, Goran PETROVSKI, Štěpán POPELKA, Yaroslav NEMESH, Miroslava SEDLÁČKOVÁ, Saskia DRUTOVIČ, Sonali ROHIWAL, Pavla JENDELOVÁ, Slaven ERCEG, Anna BRYMOVÁ, Ana ARTERO-CASTRO, Lyubomyr LYTVYNCHUK, Zbyněk STRAŇÁK, Zdeňka ELLEDEROVÁ, Jan MOTLÍK a Taras ARDAN. Advantages of nanofibrous membranes for culturing of primary RPE cells compared to commercial scaffolds. \textit{Acta Ophthalmologica}. Hoboken: Wiley, 2022, roč.~100, č.~5, s.~''E1172''-''E1185'', 14 s. ISSN~1755-375X. Dostupné z: https://dx.doi.org/10.1111/aos.15034.

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