Autor: |
Hsiao HY; Department of Biomedical Sciences, College of Medicine, Chang Gung University, Taoyuan 33302, Taiwan.; Center for Tissue Engineering, Linkuo Chang Gung Memorial Hospital, Taoyuan 33305, Taiwan., Yen TH; Department of Nephrology, Clinical Poison Center, Linkuo Chang Gung Memorial Hospital, Taoyuan 33305, Taiwan.; Department of Nephrology, College of Medicine, Chang Gung University, Taoyuan 33302, Taiwan., Wu FY; Department of Life Science, National Central University, Taoyuan 32001, Taiwan., Cheng CM; Institute of Biomedical Engineering, National Tsing Hua University, Hsinchu 300193, Taiwan., Liu JW; Center for Tissue Engineering, Linkuo Chang Gung Memorial Hospital, Taoyuan 33305, Taiwan., Fan YT; Department of Life Science, National Central University, Taoyuan 32001, Taiwan., Huang JJ; Division of Reconstructive Microsurgery, Department of Plastic and Reconstructive Surgery, Linkou Chang Gung Memorial Hospital, Taoyuan 33305, Taiwan., Nien CY; Department of Life Science, National Central University, Taoyuan 32001, Taiwan. |
Abstrakt: |
The high prevalence of kidney diseases and the low identification rate of drug nephrotoxicity in preclinical studies reinforce the need for representative yet feasible renal models. Although in vitro cell-based models utilizing renal proximal tubules are widely used for kidney research, many proximal tubule cell (PTC) lines have been indicated to be less sensitive to nephrotoxins, mainly due to altered expression of transporters under a two-dimensional culture (2D) environment. Here, we selected HK-2 cells to establish a simplified three-dimensional (3D) model using gelatin sponges as scaffolds. In addition to cell viability and morphology, we conducted a comprehensive transcriptome comparison and correlation analysis of 2D and 3D cultured HK-2 cells to native human PTCs. Our 3D model displayed stable and long-term growth with a tubule-like morphology and demonstrated a more comparable gene expression profile to native human PTCs compared to the 2D model. Many missing or low expressions of major genes involved in PTC transport and metabolic processes were restored, which is crucial for successful nephrotoxicity prediction. Consequently, we established a cost-effective yet more representative model for in vivo PTC studies and presented a comprehensive transcriptome analysis for the systematic characterization of PTC lines. |