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Cultured Human Retinal Pigment Epithelial (hRPE) Sheets: A Search for Suitable Storage Conditions

Published online by Cambridge University Press:  11 April 2018

Ayyad Z. Khan ,
Tor P. Utheim Open the ORCID record for Tor P. Utheim [Opens in a new window] ,
Sjur Reppe ,
Leiv Sandvik ,
Torstein Lyberg ,
Borghild B.-H. Roald ,
Ibrahim B. Ibrahim  and
Jon R. Eidet
Ayyad Z. Khan*
Affiliation:
Department of Medical Biochemistry, Oslo University Hospital, Kirkeveien 166, P.O. Box 4956, Nydalen, 0424 Oslo, Norway Institute of Clinical Medicine, Faculty of Medicine, University of Oslo, P.O. Box 1171, Blindern, 0318 Oslo, Norway
Tor P. Utheim
Affiliation:
Department of Medical Biochemistry, Oslo University Hospital, Kirkeveien 166, P.O. Box 4956, Nydalen, 0424 Oslo, Norway Department of Ophthalmology, Oslo University Hospital, Kirkeveien 166, P.O. Box 4956, Nydalen, 0424 Oslo, Norway Department of Ophthalmology, Drammen Hospital, Vestre Viken Hospital Trust, Dronninggata 28, P.O. Box 800, 3004 Drammen, Norway Department of Ophthalmology, Stavanger University Hospital, P.O. Box 8100, 4068 Stavanger, Norway Department of Plastic and Reconstructive Surgery, Oslo University Hospital, Kirkeveien 166, P.O. Box 4956, Nydalen, 0424 Oslo, Norway Department of Clinical Medicine, Faculty of Medicine, University of Bergen, P.O. Box 7804, 5020 Bergen, Norway Institute of Oral Biology, Faculty of Dentistry, University of Oslo, Sognsvannsveien 10, P.O. Box 1052, 0316 Oslo, Norway
Sjur Reppe
Affiliation:
Department of Medical Biochemistry, Oslo University Hospital, Kirkeveien 166, P.O. Box 4956, Nydalen, 0424 Oslo, Norway Lovisenberg Diakonale Hospital, Unger-Vetlesen Institute, P.O. Box 4970, Nydalen, 0440 Oslo, Norway
Leiv Sandvik
Affiliation:
Department of Periodontology, Institute of Clinical Odontology, Faculty of Dentistry, University of Oslo, Sognsvannsveien 10, P.O. Box 1052, 0316 Oslo, Norway
Torstein Lyberg
Affiliation:
Department of Medical Biochemistry, Oslo University Hospital, Kirkeveien 166, P.O. Box 4956, Nydalen, 0424 Oslo, Norway
Borghild B.-H. Roald
Affiliation:
Institute of Clinical Medicine, Faculty of Medicine, University of Oslo, P.O. Box 1171, Blindern, 0318 Oslo, Norway Department of Pathology, Oslo University Hospital, Kirkeveien 166, P.O. Box 4956, Nydalen, 0424 Oslo, Norway
Ibrahim B. Ibrahim
Affiliation:
Institute of Clinical Medicine, Faculty of Medicine, University of Oslo, P.O. Box 1171, Blindern, 0318 Oslo, Norway
Jon R. Eidet
Affiliation:
Department of Medical Biochemistry, Oslo University Hospital, Kirkeveien 166, P.O. Box 4956, Nydalen, 0424 Oslo, Norway Department of Ophthalmology, Oslo University Hospital, Kirkeveien 166, P.O. Box 4956, Nydalen, 0424 Oslo, Norway
*
*Author for correspondence: Ayyad Z. Khan, E-mail: [email protected]
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Abstract

The advancement of human retinal pigment epithelial cell (hRPE) replacement therapy is partly dependent on optimization of cell culture, cell preservation, and storage medium. This study was undertaken to search for a suitable storage temperature and storage medium for hRPE. hRPE monolayer sheets were cultured under standard conditions at 37°C and then randomized for storage at six temperatures (4, 16, 20, 24, 28, and 37°C) for 7 days. After revealing a suitable storage temperature, hRPE sheets were subsequently stored with and without the silk protein sericin added to the storage medium. Live/dead assay, light microscopy, pH, and phenotypic expression of various proteins were used to assess cell cultures stored at different temperatures. After 7 days of storage, hRPE morphology was best preserved at 4°C. Addition of sericin to the storage medium maintained the characteristic morphology of the preserved cells, and improved pigmentation and levels of pigmentation-related proteins in the cultured hRPE sheets following a 7-day storage period at 4°C.

Keywords

retinal pigment epitheliumcell preservationsericinregenerative medicinetissue engineering
Type
Biological Science Applications
Information
Microscopy and Microanalysis , Volume 24 , Issue 2 , April 2018 , pp. 147 - 155
Copyright
© Microscopy Society of America 2018 

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