The trisomy 16 mouse is a genetic model of Down syndrome. Clonal cell lines were developed from trisomic as well as euploid embryonic mouse septal cells by introduction of thermolabile large T antigen mutant of SV 40. The cell lines underwent morphological differentiation at the non-permissive temperature and in respone to a differentiating agent. Immunocytochemical staining indicated that cells of neuronal lineage were immortalized. The addition of β-nerve growth factor (100 ng/ml) increased the survival rate of a trisomy cell line in differentiated state, as measured by Trypan blue exclusion. These cell lines may prove useful in studies of neuronal abnormalities in this mouse model of Down syndrome.