Immortalization and Characterization of Porcine Macrophages That Had Been Transduced with Lentiviral Vectors Encoding the SV40 Large T Antigen and Porcine Telomerase Reverse Transcriptase

Takenouchi, Takato; Kitani, Hiroshi; Suzuki, Shunichi; Nakai, Michiko; Fuchimoto, Dai-ichiro; Tsukimoto, Mitsutoshi; Shinkai, Hiroki; Sato, Mitsuru; Uenishi, Hirohide

To further verify the effects of the co-expression of SV40LT and pTERT on the immortalization of porcine macrophages, we attempted to immortalize LDLR-KO pig blood-derived macrophages. When LDLR-KO pig blood-derived macrophages were infected with lentiviral particles carrying the SV40LT gene in combination with lentiviral particles carrying the pTERT gene, immortalized cells were generated and routinely passaged. The morphology of the immortalized cells closely resembled those of the IPKM and primary macrophages; i.e., they have ruffled membranes and cell processes ( Figure 6A) . The presence of the targeted genomic locus in the immortalized cells was clearly shown by PCR ( Figure 6B) . As was found for the IPKM, immunostaining demonstrated that the immortalized cells were strongly positive for macrophage markers, but negative for other cell markers (Figure 6C) . These findings suggest that inducing the co-expression of SV40LT and pTERT is an effective way of immortalizing porcine macrophages.