Silk fibers assisted long-term 3D culture of human primary urinary stem cells via inhibition of senescence-associated genes: Potential use in the assessment of chronic mitochondrial toxicity

作者全名:"Ding, Huifen; George, Sunil; Leng, Xiaoyan Iris; Ihnat, Michael; Ma, Jian-Xing; Jiang, Guochun; Margolis, David; Dumond, Julie; Zhang, Yuanyuan"

作者地址:"[Ding, Huifen; George, Sunil; Zhang, Yuanyuan] Wake Forest Univ Hlth Sci, Wake Forest Inst Regenerat Med, Winston Salem, NC 27101 USA; [Ding, Huifen] Chongqing Med Univ, Chongqing Key Lab Oral Dis & Biomed Sci, Chongqing Municipal Key Lab Oral Biomed Engn Highe, Stomatol Hosp, Chongqing, Peoples R China; [Leng, Xiaoyan Iris] Wake Forest Sch Med, Dept Biostat & Data Sci, Div Publ Hlth Sci, Winston Salem, NC USA; [Ihnat, Michael] Univ Oklahoma, Hlth Sci Ctr, Dept Pharmaceut Sci, Coll Pharm, Oklahoma City, OK USA; [Ma, Jian-Xing] Wake Forest Univ Hlth Sci, Dept Biochem, Winston Salem, NC USA; [Jiang, Guochun; Margolis, David] Univ N Carolina, HIV Cure Ctr, Chapel Hill, NC USA; [Dumond, Julie] Univ N Carolina, UNC Eshelman Sch Pharm, Div Pharmacotherapy & Expt Therapeut, Chapel Hill, NC USA"

通信作者:"Zhang, YY (通讯作者),Wake Forest Univ Hlth Sci, Wake Forest Inst Regenerat Med, Winston Salem, NC 27101 USA."

来源:MATERIALS TODAY ADVANCES

ESI学科分类: 

WOS号:WOS:000827236100005

JCR分区:Q1

影响因子:10

年份:2022

卷号:15

期号: 

开始页: 

结束页: 

文献类型:Article

关键词:Mitochondrial toxicity; Antiviral drugs; 3D cellular assay; Human stem cells; Silk fibers; Spheroids

摘要:"Despite being widely applied in drug development, existing in vitro 2D cell-based models are not suitable to assess chronic mitochondrial toxicity. A novel in vitro assay system mimicking in vivo micro -environment for this purpose is urgently needed. The goal of this study is to establish a 3D cell plat-form as a reliable, sensitive, cost-efficient, and high-throughput assay to predict drug-induced mito-chondrial toxicity. We evaluated a long-term culture of human primary urine-derived stem cells (USC) seeded in 3D silk fiber matrix (3D USC-SFM) and further tested chronic mitochondrial toxicity induced by Zalcitabine (ddC, a nucleoside reverse transcriptase inhibitor) as a test drug, compared to USC grown in spheroids. The numbers of USC remain steady in 3D spheroids for 4 weeks and 3D SFM for 6 weeks. However, the majority (95%) of USC survived in 3D SFM, while cell numbers significantly declined in 3D spheroids at 6 weeks. Highly porous SFM provides large-scale numbers of cells by increasing the yield of USC 125-fold/well, which enables the carrying of sufficient cells for multiple experiments with less labor and lower cost, compared to 3D spheroids. The levels of mtDNA content and mitochondrial superoxide dismutase2 [SOD2] as an oxidative stress biomarker and cell senescence genes (RB and P16, p21) of USC were all stably retained in 3D USC-SFM, while those were significantly increased in spheroids. mtDNA content and mitochondrial mass in both 3D culture models significantly decreased six weeks after treatment of ddC (0.2, 2, and 10 mM), compared to 0.1% DMSO control. Levels of complexes I, II, and III significantly decreased in 3D SFM-USC treated with ddC, compared to only complex I level which declined in spheroids. A dose-and time-dependent chronic MtT displayed in the 3D USC-SFM model, but not in spheroids. Thus, a long-term 3D culture model of human primary USC provides a cost-effective and sensitive approach potential for the assessment of drug-induced chronic mitochondrial toxicity. (C) 2022 The Authors. Published by Elsevier Ltd."

基金机构:"National Institute of Allergy and Infectious Diseases, National Institutes of Health [R21 AI152832, R03 AI165170]"

基金资助正文:"We thank Ms. Lilly M. Wong for her editing. This project has been funded with Federal funds from the National Institute of Allergy and Infectious Diseases, National Institutes of Health, under Contract No. R21 AI152832 and R03 AI165170. (PI: Y. Z) ."