Development and Characterization of an Immortal and Differentiated Murine Trabecular Meshwork Cell Line

METHODS. Mouse TM in situ was studied by light and electron microscopy (EM). In addition, TM was isolated from the H-2K-tsA58 transgenic mouse strain in which promoter sequences of the major histocompatibility complex H-2K class 1 gene are fused to sequences of the SV40 mutant temperature-sensitive (ts) strain tsA58. The promoter is inducible by interferon (IFN)-y, and the tsA58 gene product is active at 33°C (permissive conditions), but not at 37°C (nonpermissive conditions). The TM explant was cultured in permissive conditions. Outgrowing cells were passaged through two rounds of single-cell cloning. One clonal cell line (MUTM-NEI/1) was characterized in nonpermissive conditions by EM, immunohistochemistry, reverse transcriptionpolymerase chain reaction (RT-PCR), and northern blot hybridization. In addition, MUTM-NEI/1 cells were transfected with plasmid DNA.