Department of Immunology, Kyoto Prefecture University of Medicine, Kamigyo-ku, Kyoto, 602-8566, Japan.
Department of Urology, Kyoto Prefecture University of Medicine, Kamigyo-ku, Kyoto, 602-8566, Japan.
Sci Rep. 2019 Sep 25;9(1):13850. doi: 10.1038/s41598-019-50388-6.
Urothelial cells play essential roles in protection of urine exudation and bacterial invasion at the urothelial mucosa, so that defect or damage of urothelial cells associated with urinary tract diseases may cause serious problems. If a sufficient number of functional urothelial cells are prepared in culture and transplanted into the damaged urothelial lesions, such technology may provide beneficial effects to patients with diseases of the urinary tract. Here we found that human adult dermal fibroblasts were converted into urothelial cells by transducing genes for four transcription factors, FOXA1, TP63, MYCL and KLF4 (FTLK). The directly converted urothelial cells (dUCs) formed cobblestone-like colonies and expressed urothelium-specific markers. dUCs were successfully expanded and enriched after serial passages using a specific medium that we optimized for the cells. The passaged dUCs showed similar genome-wide gene expression profiles to normal urothelial cells and had a barrier function. The FTLK-transduced fibroblasts were also converted into urothelial cells in vivo and recruited to the regenerating urothelial tissue after they were transplanted into the bladder of mice with interstitial cystitis. Our technology may provide a promising solution for a number of patients with urinary tract disorders.
尿路上皮细胞在保护尿路上皮黏膜免受尿液渗出和细菌侵袭方面发挥着重要作用,因此与泌尿道疾病相关的尿路上皮细胞的缺陷或损伤可能会导致严重的问题。如果在培养中制备足够数量的功能性尿路上皮细胞并将其移植到受损的尿路上皮病变部位,这种技术可能会为患有泌尿道疾病的患者带来有益的效果。在这里,我们发现通过转导四个转录因子(FOXA1、TP63、MYCL 和 KLF4)的基因,人成体皮肤成纤维细胞可以转化为尿路上皮细胞。直接转化的尿路上皮细胞(dUC)形成鹅卵石样集落,并表达尿路上皮特异性标志物。我们优化了一种专门用于这些细胞的培养基,通过连续传代成功地扩增和富集了 dUC。传代的 dUC 显示出与正常尿路上皮细胞相似的全基因组基因表达谱,并具有屏障功能。FTLK 转导的成纤维细胞也在体内转化为尿路上皮细胞,并在将其移植到患有间质性膀胱炎的小鼠膀胱后被募集到再生的尿路上皮组织中。我们的技术可能为许多患有泌尿道疾病的患者提供了一种有前途的解决方案。