Effect of Storage Temperature on Structure and Function of Cultured Human Oral Keratinocytes.
To assess the effect of storage temperature on the viability, phenotype, metabolism, and morphology of cultured human oral keratinocytes (HOK).Cultured HOK cells were stored in HEPES- and sodium bicarbonate-buffered Minimum Essential Medium (MEM) at nine temperatures in approximately 4 °C increments...
| Main Authors: | , , , , , , , , |
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| Format: | Article |
| Language: | English |
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Public Library of Science (PLoS)
2015-01-01
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| Series: | PLoS ONE |
| Online Access: | http://europepmc.org/articles/PMC4459984?pdf=render |
| _version_ | 1811296321380810752 |
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| author | Rakibul Islam Catherine Jackson Jon R Eidet Edward B Messelt Rima Maria Corraya Torstein Lyberg May Griffith Darlene A Dartt Tor P Utheim |
| author_facet | Rakibul Islam Catherine Jackson Jon R Eidet Edward B Messelt Rima Maria Corraya Torstein Lyberg May Griffith Darlene A Dartt Tor P Utheim |
| author_sort | Rakibul Islam |
| collection | DOAJ |
| description | To assess the effect of storage temperature on the viability, phenotype, metabolism, and morphology of cultured human oral keratinocytes (HOK).Cultured HOK cells were stored in HEPES- and sodium bicarbonate-buffered Minimum Essential Medium (MEM) at nine temperatures in approximately 4 °C increments from 4 °C to 37 °C for seven days. Cells were characterized for viability by calcein fluorescence, phenotype retention by immunocytochemistry, metabolic parameters (pH, glucose, lactate, and O2) within the storage medium by blood gas analysis, and morphology by scanning electron microscopy and light microscopy.Relative to the cultured, but non-stored control cells, a high percentage of viable cells were retained only in the 12 °C and 16 °C storage groups (85% ± 13% and 68% ± 10%, respectively). Expression of ABCG2, Bmi1, C/EBPδ, PCNA, cytokeratin 18, and caspase-3 were preserved after storage in the 5 groups between 4 °C and 20 °C, compared to the non-stored control. Glucose, pH and pO2 in the storage medium declined, whereas lactate increased with increasing storage temperature. Morphology was best preserved following storage of the three groups between 12 °C, 16 °C, and 20 °C.We conclude that storage temperatures of 12 °C and 16 °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. |
| first_indexed | 2024-04-13T05:46:35Z |
| format | Article |
| id | doaj.art-5e768acf84e849768b3a8619f21edd5e |
| institution | Directory Open Access Journal |
| issn | 1932-6203 |
| language | English |
| last_indexed | 2024-04-13T05:46:35Z |
| publishDate | 2015-01-01 |
| publisher | Public Library of Science (PLoS) |
| record_format | Article |
| series | PLoS ONE |
| spelling | doaj.art-5e768acf84e849768b3a8619f21edd5e2022-12-22T02:59:56ZengPublic Library of Science (PLoS)PLoS ONE1932-62032015-01-01106e012830610.1371/journal.pone.0128306Effect of Storage Temperature on Structure and Function of Cultured Human Oral Keratinocytes.Rakibul IslamCatherine JacksonJon R EidetEdward B MesseltRima Maria CorrayaTorstein LybergMay GriffithDarlene A DarttTor P UtheimTo assess the effect of storage temperature on the viability, phenotype, metabolism, and morphology of cultured human oral keratinocytes (HOK).Cultured HOK cells were stored in HEPES- and sodium bicarbonate-buffered Minimum Essential Medium (MEM) at nine temperatures in approximately 4 °C increments from 4 °C to 37 °C for seven days. Cells were characterized for viability by calcein fluorescence, phenotype retention by immunocytochemistry, metabolic parameters (pH, glucose, lactate, and O2) within the storage medium by blood gas analysis, and morphology by scanning electron microscopy and light microscopy.Relative to the cultured, but non-stored control cells, a high percentage of viable cells were retained only in the 12 °C and 16 °C storage groups (85% ± 13% and 68% ± 10%, respectively). Expression of ABCG2, Bmi1, C/EBPδ, PCNA, cytokeratin 18, and caspase-3 were preserved after storage in the 5 groups between 4 °C and 20 °C, compared to the non-stored control. Glucose, pH and pO2 in the storage medium declined, whereas lactate increased with increasing storage temperature. Morphology was best preserved following storage of the three groups between 12 °C, 16 °C, and 20 °C.We conclude that storage temperatures of 12 °C and 16 °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.http://europepmc.org/articles/PMC4459984?pdf=render |
| spellingShingle | Rakibul Islam Catherine Jackson Jon R Eidet Edward B Messelt Rima Maria Corraya Torstein Lyberg May Griffith Darlene A Dartt Tor P Utheim Effect of Storage Temperature on Structure and Function of Cultured Human Oral Keratinocytes. PLoS ONE |
| title | Effect of Storage Temperature on Structure and Function of Cultured Human Oral Keratinocytes. |
| title_full | Effect of Storage Temperature on Structure and Function of Cultured Human Oral Keratinocytes. |
| title_fullStr | Effect of Storage Temperature on Structure and Function of Cultured Human Oral Keratinocytes. |
| title_full_unstemmed | Effect of Storage Temperature on Structure and Function of Cultured Human Oral Keratinocytes. |
| title_short | Effect of Storage Temperature on Structure and Function of Cultured Human Oral Keratinocytes. |
| title_sort | effect of storage temperature on structure and function of cultured human oral keratinocytes |
| url | http://europepmc.org/articles/PMC4459984?pdf=render |
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