Fig. 2
Isolation and immortalization of SMGCs (iSMGCs) and SLGCs (iSLGCs). A Primary SMGCs or SLGCs were isolated on P0 SMG or SLG (captured under transmission light) and cultured in complete DMEM medium. a Morphology of primary cells at 24 h after planting, in addition to passage 3 (p2) and p4. b Establishment iSMGCs and iSLGCs. Primary SMGCs or SLGCs were infected with retrovirus SSR#41 and selected in hygromycin-containing medium for 3 days. The surviving cells were designated as iSMGCs or iSLGCs. The immortalized cells of iSMGCs or iSLGCs were photographed at the indicated time and maintained indefinitely (passage 25 is shown). B Cell proliferation was assessed via crystal violet staining assay. The same number of primary SMGCs or SLGCs (passage 2) and iSLGCs (a1) or iSMGCs (a2) were seeded at a low density and fixed with paraformaldehyde for crystal violet staining at the indicated timepoints. The stained cells of iSMGCs (b1) and iSLGCs (b2) were dissolved in 10% acetic acid and detached for OD measurement to quantitatively determine at A570 nm to A590 nm. The assays were performed in triplicate. **P < 0.01