We investigated chromosome (Chr) aberrations in the process of replicative senescence and immortalization of cultured bovine oviduct epithelial cells (BOEC) before and after transfecting vectors SV40 large T or human telomerase reverse transcriptase (hTERT). We found that a gradual increase in the number of metacentric chromosomes occurred during replicative senescence but not immortalization of BOEC. The accumulation of metacentric chromosomes was concomitant with decreases in the number of acrocentric autosomes, strongly suggesting that Robertsonian (Rb) translocation frequently occurred in cultured BOEC. The process was also correlated with an accumulation of extremely shortened telomeres (<4 kb). The maximum number of metacentric chromosomes reached a plateau (8.75±0.53) in the senescent BOEC (approximately 48 population doublings), and the value was stably maintained in all immortalized lines. These results suggest that not all autosomes may be involved in Rb translocation. Fluorescence in situ hybridization analysis using probes specific for Chr1, Chr29, telomeres, and x-chromosomes of bovine confirmed the presence of t(1;29) with other unidentified fused chromosomes. There was no evidence for duplication of sex chromosomes. Because no detectable fluorescence in situ hybridization signals at the centromere for telomeres were indicative of no direct integration of telomere sequences in the Rb translocated chromosomes, these results raise a possibility that Rb translocation between certain autosomes of bovine cells is partly but critically dependent upon a physical state of telomere attrition. The cells and cell lines established in this study could provide a promising system for further studies on the mechanisms of chromosomal translocation because of centromeric fusion in bovine cells.