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Establishment of primary cell culture from stria vascularis explants. Morphological and functional characterization.

Authors
Kim, HN; Chang, MS; Chung, MH; Park, K
Citation
Acta oto-laryngologica, 116(6):805-811, 1996
Journal Title
Acta oto-laryngologica
ISSN
0001-64891651-2251
Abstract
To provide the prerequisite for long-term study of the inner ear related to structural and functional integrity, tissue of stria vascularis with spiral ligament was isolated from Wistar rat cochleas and cultured using the explant-culture technique. The following culture media were used: EMEM with Hepes buffer, hydrocortisone (400 ng/ml), transferrin (5 micrograms/ml). triiodothyronine (10(-9) M), cholera toxin (10(-10) M), insulin (5 micrograms/ml), and epidermal growth factor (10 ng/ml). To characterize the cells growing out from the explant, immunofluorescence with cytokeratin (cytokeratin 18) and ultrastructural examination with SEM and TEM were performed. The marginal cell function was investigated by expression of Na+, K(+)-ATPase antisera against beta 2 subunit of rat Na+, K(+)-ATPase and P-NPPase. We were able to maintain the cultured cells for 3 weeks or more. Monolayered marginal cells were observed beyond 14 days in vitro and the expression of cytokeratin 18 was especially enhanced. The cultured marginal cells were almost identical to in vivo cells both as regards ultrastructural features and Na+, K(+)-ATPase activity. The present results suggest that the primary explant culture technique is a reliable in vitro model of strial marginal cells. However, establishment of the cell line is needed for long-term study.
MeSH terms
AnimalsCell DivisionCells, CulturedCochlea/*ultrastructureImmunohistochemistryKeratinsMicroscopy, ElectronRatsRats, WistarSodium-Potassium-Exchanging ATPase/ultrastructureStria Vascularis/*ultrastructure
PMID
8973711
Appears in Collections:
Journal Papers > School of Medicine / Graduate School of Medicine > Otolaryngology
AJOU Authors
박, 기현
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