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The effect of human serum albumin on the extended storage of human oral keratinocyte viability under mild hypothermia

 Eun Ju Lee  ;  Seung-Ae Lee  ;  Jin Kim 
 CRYOBIOLOGY, Vol.50(1) : 103-111, 2005 
Journal Title
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Antioxidants/pharmacology ; Apoptosis ; Cell Cycle ; Cell Survival ; Cells, Cultured ; Collagen Type I/metabolism ; Culture Media/metabolism ; Culture Media/pharmacology* ; Fibroblasts/metabolism ; Flow Cytometry ; Gingiva/metabolism ; Humans ; Hypothermia, Induced ; Keratinocytes/cytology* ; Keratinocytes/metabolism ; Organ Preservation Solutions/pharmacology ; Serum Albumin/chemistry* ; Sodium Chloride/pharmacology ; Temperature ; Time Factors ; Tissue Preservation/methods
Primary human oral keratinocytes ; Preservation solutions ; Human serum albumin ; Hypothermic condition
Isolated oral keratinocytes in suspension provide a number of advantages for use in maxillofacial surgery, however, the poor stability of this cell preparation at physiological temperatures is an apparent barrier preventing their use. The purpose of the present study was to evaluate whether human serum albumin (HSA) could serve as an effective constituent of a storage medium to enhance human oral keratinocyte (HOK) viability under conditions of mild hypothermia. Primary human oral keratinocytes were isolated from small pieces of the non-inflamed gingival tissues obtained during the extraction of the third molars of patients. HOK were cultured on collagen type I-coated culture dishes in keratinocyte growth medium (KGM). After the trypsinization of a culture dish (passage 2 or 3), freshly isolated HOK were stored for 24, 48, and 72 h at 4 °C or at room temperature in KGM, saline, Dulbecco’s modified Eagle’s medium (DMEM), saline supplemented with 10% HSA or DMEM supplemented with 10% (v/v) HSA under one atmosphere pressure. After storage, HOK cell survival was determined by dye exclusion using trypan blue and colony-forming assay and cell cycle change was obtained by flow cytometry. Highest cell viability was obtained in saline supplemented with 10% HSA and DMEM supplemented with 10% (v/v) HSA at 4 °C and at room temperature. Under these conditions no significant decline in keratinocyte viability was observed for at least 48 h. The cell cycle profiles of these cells were also maintained for at least 48 h at room temperature. These observations demonstrate that HSA might be better at preserving the viability of HOK stored under hypothermic and mild hypothermic conditions up to 48 h.
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2. College of Dentistry (치과대학) > Dept. of Oral Pathology (구강병리학교실) > 1. Journal Papers
Yonsei Authors
Kim, Jin(김진)
Lee, Seung Ae(이승애)
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