抄録
Freshly isolated bovine oviduct epithelial cells (BOECs) were processed to long-term cultivation until the cell multiplication potential is limited in vitro. Replicative senescence is monitored by the expression of senescence-associated β-galactosidase and a significant reduction in sizes of telomere restriction fragment (TRF). Immortalized cell lines were isolated after transfecting SV40 large T or hTERT. Several independently isolated BOEC lines; BO+SV (a, b, c and d) and BO+TERT (a and b) were established as a clonal cell line that harbor SV40large T or activated hTERT. Since BO+SV lines maintained TRF without telomerase activation, these lines achieved immortality due to ALT (alternative lengthening of telomeres). In contrast, BO+TERT lines show progressive telomere elongation along with the strong telomerase activity. These results demonstrate that the immortalized BOEC lines by the oncogenic virus SV40 large T or telomerase catalytic subunit hTERT generated distinctive cell lines in which telomere erosion was protected by either ALT or the activated telomerase, respectively. Karyotype analysis showed that all immortal BOEC lines had a number of metacentric chromosomes resulted from acrocentric chromosome fusion. These BOECs lines may be useful for investigating various aspects of cytogenetics and cell genetics in terms of telomere dynamics and genomic instability.
