Immortalization of Urine-Derived UM51-PrePodo Cells

The cell line UM51-PrePodo was derived from the urine of a 51-year-old male of African origin. The cells were cultured, and differentiation was induced as described in [41 (link)]. Immortalization of the cells was achieved by cationic polymer transfection of the pCDNA-3xHA-hTERT plasmid with Xfect (Takara BIO INC, Kusatsu, Shiga Prefecture, Japan). The plasmid pCDNA-3xHA-hTERT was obtained from (Addgene plasmid # 51637; http://n2t.net/addgene:51637; RRID: Addgene_51637; accessed 12 January 2023) [42 (link)]. In brief, 2 µg of the plasmid was incubated at RT for 10 min with 100 µL of transfection buffer and 1 µL of transfection reagent. After incubation, the mix was added to at least 50% confluent 6 well of UM51-PrePodo, growing as a monolayer. In another transfection, 2 µg of the commercial pmaxGFP Vector (Lonza, Basel, Switzerland) was incubated at RT for 10 min with 100 µL of transfection buffer and 1 µL of transfection reagent. After incubation, the mix was added to at least 50% confluent 6 well of UM51-PrePodo-hTERT, growing as monolayer.