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.

Basic information

Original name

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

Authors

TICHOTOVÁ, Lucie (203 Czech Republic), Hana STUDENOVSKÁ (203 Czech Republic), Goran PETROVSKI, Štěpán POPELKA (203 Czech Republic), Yaroslav NEMESH, Miroslava SEDLÁČKOVÁ (203 Czech Republic, belonging to the institution), Saskia DRUTOVIČ, Sonali ROHIWAL, Pavla JENDELOVÁ (203 Czech Republic), Slaven ERCEG, Anna BRYMOVÁ (203 Czech Republic), Ana ARTERO-CASTRO, Lyubomyr LYTVYNCHUK, Zbyněk STRAŇÁK (203 Czech Republic), Zdeňka ELLEDEROVÁ, Jan MOTLÍK and Taras ARDAN (guarantor)

Edition

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

Other information

Language

English

Type of outcome

Článek v odborném periodiku

Field of Study

30207 Ophthalmology

Country of publisher

United States of America

Confidentiality degree

není předmětem státního či obchodního tajemství

References:

URL

Impact factor

Impact factor: 3.400

RIV identification code

RIV/00216224:14110/22:00125041

Organization unit

Faculty of Medicine

DOI

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

UT WoS

000710006200001

Keywords in English

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

Abstract

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 and Taras ARDAN. Advantages of nanofibrous membranes for culturing of primary RPE cells compared to commercial scaffolds. Acta Ophthalmologica. Hoboken: Wiley, 2022, vol. 100, No 5, p. "E1172"-"E1185", 14 pp. ISSN 1755-375X. Available from: 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 and Taras ARDAN. Advantages of nanofibrous membranes for culturing of primary RPE cells compared to commercial scaffolds. \textit{Acta Ophthalmologica}. Hoboken: Wiley, 2022, vol.~100, No~5, p.~''E1172''-''E1185'', 14 pp. ISSN~1755-375X. Available from: https://dx.doi.org/10.1111/aos.15034.

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