Effect of Storage Temperature on Structure and Function of Cultured Human Oral Keratinocytes

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Islam, RakibulUniversity of Oslo, Norway; Oslo University Hospital, Norway; Harvard University, MA USA (author)

Effect of Storage Temperature on Structure and Function of Cultured Human Oral Keratinocytes

Article/chapterEnglish2015

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2015-06-08
Public Library of Science,2015
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LIBRIS-ID:oai:DiVA.org:liu-120047
https://urn.kb.se/resolve?urn=urn:nbn:se:liu:diva-120047URI
https://doi.org/10.1371/journal.pone.0128306DOI

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Language:English
Summary in:English

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Funding Agencies|Department of Oral Biology, Faculty of Dentistry, University of Oslo, Oslo, Norway [5142001]
Purpose/Aims To assess the effect of storage temperature on the viability, phenotype, metabolism, and morphology of cultured human oral keratinocytes (HOK). Materials and Methods Cultured HOK cells were stored in HEPES- and sodium bicarbonate-buffered Minimum Essential Medium (MEM) at nine temperatures in approximately 4 degrees C increments from 4 degrees C to 37 degrees C for seven days. Cells were characterized for viability by calcein fluorescence, phenotype retention by immunocytochemistry, metabolic parameters (pH, glucose, lactate, and O-2) within the storage medium by blood gas analysis, and morphology by scanning electron microscopy and light microscopy. Results Relative to the cultured, but non-stored control cells, a high percentage of viable cells were retained only in the 12 degrees C and 16 degrees C storage groups (85%+/- 13% and 68%+/- 10%, respectively). Expression of ABCG2, Bmi1, C/EBP delta, PCNA, cytokeratin 18, and caspase-3 were preserved after storage in the 5 groups between 4 degrees C and 20 degrees C, compared to the non-stored control. Glucose, pH and pO(2) in the storage medium declined, whereas lactate increased with increasing storage temperature. Morphology was best preserved following storage of the three groups between 12 degrees C, 16 degrees C, and 20 degrees C. Conclusion We conclude that storage temperatures of 12 degrees C and 16 degrees C were optimal for maintenance of cell viability, phenotype, and morphology of cultured HOK. The storage method described in the present study may be applicable for other cell types and tissues; thus its significance may extend beyond HOK and the field of ophthalmology.

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MEDICIN OCH HÄLSOVETENSKAP Klinisk medicin hsv//swe
MEDICAL AND HEALTH SCIENCES Clinical Medicine hsv//eng

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Jackson, CatherineOslo University Hospital, Norway (author)
Eidet, Jon R.Oslo University Hospital, Norway (author)
Messelt, Edward B.University of Oslo, Norway (author)
Maria Corraya, RimaOslo University Hospital, Norway; Harvard University, MA USA (author)
Lyberg, TorsteinOslo University Hospital, Norway (author)
Griffith, MayLinköpings universitet,Avdelningen för cellbiologi,Medicinska fakulteten(Swepub:liu)maygr76 (author)
Dartt, Darlene A.Harvard University, MA USA (author)
Utheim, Tor P.University of Oslo, Norway; Oslo University Hospital, Norway; Harvard University, MA USA (author)
University of Oslo, Norway; Oslo University Hospital, Norway; Harvard University, MA USAOslo University Hospital, Norway (creator_code:org_t)

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In:PLOS ONE: Public Library of Science10:6, s. e0128306-1932-6203

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