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Cellosaurus publication CLPUB00246

Publication number CLPUB00246
Authors Belyantseva I.A., Kalinec G.M., Kalinec F., Kachar B.
Title In vitro differentiation of two immortalized cell lines derived from the stria vascularis of a transgenic mouse.
Citation (In conference) Proceedings of the Association for Research in Otolaryngology 21st midwinter meeting; pp.A620-A620; Association for Research in Otolaryngology; St. Petersburg Beach; USA (1998)
Abstract We previously have established several inner ear cell lines from a two- week old transgenic mouse (Immortomouse) harboring the thermolabile immortalizing gene tsA58. These cell lines are capable of rapid proliferation when cultured at 33 C in the presence of interferon-gamma (IFN) and have the potential to differentiate when cultured at 39 C in the absence of IFN (non-permissive condition). When cultured in the DMEM medium supplemented with 10% fetal bovine serum (FBS), these cell lines are able to express the inner ear specific protein OCP-2 but they neither fully differentiate nor exhibit the structural characteristics of the cells from which they are derived. For the present study, we selected two of these cell lines, SVK-1 and SVK-2, which were derived from the stria vascularis. We cultured them in the presence of the growth factors TGF- alpha, IGF1, EGF, FGF2, BDNF, NT3, which were applied individually or in combination in order to determine whether they could induce the differentiation process. Cultures were monitored by phase contrast light microscopy and by immunofluorescence using antibodies against Nestin, a marker for neuroepithelial stem-like cells and against occludin, a tight junction protein used as a marker for differentiated epithelial cells. The cells initially were allowed to divide at the permissive condition (33 C + IFN) and then were maintained in a medium with 5% FBS without IFN for one week prior to culturing at 39 C. At the permissive condition the cells were pleomorphic and expressed nestin, but during the first week at the non-permissive condition they began to show contact inhibition, great reduction in nestin expression, and exhibited an epithelial-like shape. Addition of growth factors stimulated the cells to further differentiate. EGF (100 ng/ml) was the most effective factor for the SVK-1, while the combination of TGF-alpha (100 ng/ml) and IGF1 (100 ng/ml) was the most effective for the SVK-2. Under these conditions both cell lines formed tight junctions and organized into a regular polygonal epithelial monolayer that resembled the marginal or basal cells of the stria vascularis. Thus, these new cell lines probably represent true stria vascularis epithelial cells, and maybe useful for future studies of stria vascularis function.
Cell lines CVCL_D805; SVK-1
CVCL_D806; SVK-2