||Human Homo sapiens
||Shi Q, King RW. Chromosome nondisjunction yields tetraploid rather than aneuploid cells in human cell lines. Nature. 2005 Oct 13 437(7061):1038-42. Supplementary online material p. 4PubMed ID16222248
obtain H2B-GFP positive N/TERT-1 cells, a retroviral vector expressing an H2B-GFP
fusion protein was constructed by replacing the EGFP fragment in retroviral vector
pLEGFP-N1 (BD Clontech 6059-1) with the fragment H2BGFP from pBOS-H2BGFP
(BD Pharmingen 559241) cut out with Not I and Sal I. To obtain cell lines that stably express H2B-GFP, N/TERT-1 cells were grown
on 6-well plates (2×10^5 per well) for 48 h and transduced by refeeding for 10 h with
retroviral supernatant plus 2 µg of Polybrene (Sigma) per ml. The transduced cells were
subcultured the next day into 75-cm^2 flasks. Drug selection (200 µg/ml G418) was started
2 days after transduction and continued for 2 weeks.
||N/TERT-1 cells found to have a normal male karyotype.