Production of Conditionally Immortalised Cell Lines from a Transgenic Mouse

Abstract

This review describes the H2k^btsA58 transgenic mouse (Immortomouse) and its application to the production of conditionally immortalised cell lines from sensory epithelia within the mammalian inner ear. Established cell lines should overcome many of the technical difficulties associated with experimental procedures in auditory and vestibular research. These include the limited amount of tissue available and the relatively complex and laborious dissection. Conditional immortalisation should also allow essential studies on the molecular and cellular mechanisms that govern both the differentiation of sensory cells and the development of sensory epithelia.

A Cochlear Cell Line as an in vitro System for Drug Ototoxicity Screening

“…Establishment of immortal cell lines has been facilitated by the development of a transgenic mouse, Immortomouse™, which harbors a temperature-sensitive mutant of the SV40 large T antigen gene under the control of an interferon-Á-inducible promoter element [Jat et al, 1991;Holley and Lawlor, 1997]. Several cochlear and vestibular cell lines have already been developed from the Immortomouse [Barald et al, 1997;Rivolta et al, 1998;Kalinec et al, 1999;Lawlor et al, 1999], and are currently being used in a variety of research projects (e.g.…”

Gentamicin-Induced Cytotoxicity Involves Protein Kinase C Activation, Glutathione Extrusion and Malondialdehyde Production in an Immortalized Cell Line from the Organ of Corti

“…In recent years, several immortalized cell lines derived from the mouse or rat vestibule and organ of Corti have been developed and characterized [Barald et al, 1997;Holley and Lawlor, 1997;Lawlor et al, 1999;Rivolta et al, 1998;Zheng et al, 1998]. OC-k3 cells, derived from the organ of Corti of transgenic mice, were utilized in the present study [Kalinec et al, 1999].…”

Auditory hair cell precursors immortalized from the mammalian inner ear

“…Conditionally immortal cell lines from mammalian auditory sensory epithelia should provide important experimental tools in the study of the development, function and regeneration of hearing (Barald et al 1997;Holley & Lawlor 1997). The mammalian cochlea possesses just a few thousand sensory hair cells (Echteler et al 1994), for which there is no demonstrable regeneration after embryonic development (Kelley et al 1995;Corwin & Oberholtzer 1997;Zine & Ribaupierre 1998).…”

“…Constrained by cell numbers and the absence of cell division in vitro, we aimed to derive conditionally immortalized hair cell progenitors from the H-2Kb-tsA58 transgenic mouse (Jat et al 1991;Holley & Lawlor 1997). This mouse carries a conditionally expressed, temperature-sensitive immortalizing gene that perpetuates cell division, preventing terminal di¡erentiation.…”

“…In the mouse, most cochlear hair cells pass through this stage by E14 (Ruben 1967;Kelley et al 1993) and there is no evidence that precursors or stem cells remain in the cochlea after that time. Thus we decided to select sensory epithelial cells from tissue at stage E13 (Holley & Lawlor 1997). The aim was to immortalize sensory cell precursors before they were fully di¡eren-tiated, minimizing possible artefacts caused by having to dedi¡erentiate and redi¡erentiate a mature cell, thus establishing a cell line that would resemble as closely as possible the normal process of development and di¡eren-tiation.…”