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Background Prostate cancer cell lines from diverse backgrounds are believed to hold the key to addressing current disparities in prostate cancer incidence and mortality rates among Black men. Despite decades of established cell line protocols, developing new cell lines, especially from the prostate, continues to attract less than ideal success rates (<10%). Methods ACRJ-PC28 was developed from a transrectal needle biopsy (TRNB) that was mechanically dissociated and grown on 3T3 STO fibroblast feeder cells followed by establishment via inactivation of the CDKN2A locus and simultaneous expression of human telomerase. Characterization assays were carried out using growth curve analysis, immunoblots, immunohistochemistry, 3D cultures, immunofluorescence imaging, confocal microscopy, WGS and, RNA-Seq. Results ACRJ-PC28 has been passaged more than 40 times in vitro over 10 months with a …