Improving in vitro Sertoli cell/gonocyte co-culture model for assessing male reproductive toxicity: Lessons learned from comparisons of cytotoxicity versus genomic responses to phthalates.
Improving in vitro Sertoli cell/gonocyte co-culture model for assessing male reproductive toxicity: Lessons learned from comparisons of cytotoxicity versus genomic responses to phthalates.
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DOI：
10.1016/j.taap.2009.06.014
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发表时间：
2009-09-15
影响因子：
3.8
通讯作者：
Faustman EM
中科院分区：
文献类型：
--
作者：
Yu X;Hong S;Moreira EG;Faustman EM
性腺细胞存在于新生儿睾丸中，是一种暂时性的雄性生殖系干细胞。研究表明，干细胞的自我更新和后代的产生可能是由体内邻近的分化细胞和细胞外基质（ECM）控制的。最近，我们开发了一种体外三维（3D）支持细胞/性腺细胞共培养（SGC）模型，该模型具有ECM覆盖层，可创建体内样生态位，并支持生殖系干细胞在3D环境中发挥功能。在本研究中，我们应用形态学和细胞毒性评估，以及基于微阵列的基因表达来检测不同邻苯二甲酸酯（PE）对该模型的影响。对体内已知雄性发育毒性pe （DTPE）和发育无毒pe （DNTPE）进行了评价。我们观察到，与DNTPE治疗的小鼠相比，DTPE诱导的剂量依赖性形态学改变、细胞活力下降和细胞毒性增加显著增加。此外，与DNTPE相比，DTPE对基因表达的影响更大，非监督聚类分析可以区分DTPE和DNTPE。我们基于系统的GO-Quant分析显示，DTPE治疗后，参与细胞周期、磷酸盐运输和凋亡调控的基因通路发生了显著变化，而DNTPE治疗后则没有。在DTPE组中观察到类固醇生成相关基因如Star、Cyp19a1、Hsd17b8和Nr4a3的表达中断，而在DNTPE组中没有。总之，我们对PEs诱导的细胞活力、细胞毒性和基于微阵列的基因表达分析的观察表明，我们的体外3D-SGC系统模拟了PEs在体内的反应，并表明3D-SGC系统可能有助于识别发育性生殖毒物。
Gonocytes exist in the neonatal testis and represent a transient population of male germ-line stem cells. It has been shown that stem cell self-renewal and progeny production is probably controlled by the neighboring differentiated cells and extracellular matrix (ECM) in vivo known as niches. Recently, we developed an in vitro three-dimensional (3D) Sertoli cell/Gonocyte coculture (SGC) model with ECM overlay, which creates an in vivo-like niche and supports germ-line stem cell functioning within a 3D environment. In this study, we applied morphological and cytotoxicity evaluations, as well as microarray-based gene expression to examine the effects of different phthalate esters (PE) on this model. Known in vivo male developmentally toxic PEs (DTPE) and developmentally nontoxic PEs (DNTPE) were evaluated. We observed that DTPE induced significantly greater dose-dependent morphological changes, a decrease in cell viability and an increase in cytotoxicity compared to those treated with DNTPE. Moreover, gene expression was more greatly altered by DTPE than by DNTPE and non-supervised cluster analysis allowed the discrimination of DTPE from the DNTPE. Our systems-based GO-Quant analysis showed significant alterations in gene pathways involved in cell cycle, phosphate transport and apoptosis regulation with DTPE but not with DNTPE treatment. Disruptions of steroidogenesis related-gene expression such as Star, Cyp19a1, Hsd17b8, and Nr4a3 were observed in the DTPE group, but not in the DNTPE group. In summary, our observation on cell viability, cytotoxicity, and microarray-based gene expression analysis induced by PEs demonstrate that our in vitro 3D-SGC system mimicked in vivo responses for PEs and suggests that the 3D-SGC system might be useful in identifying developmental reproductive toxicants.
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影响因子：
3.8
作者：
Hamadeh, HK;Bushel, PR;Afshari, CA
通讯作者：
Afshari, CA
影响因子：
3.8
作者：
Lampen, A;Zimnik, S;Nau, H
通讯作者：
Nau, H
影响因子：
14.9
作者：
Harris, MA;Clark, J;White, R
通讯作者：
White, R
影响因子：
3.8
作者：
CHAPIN, RE;GRAY, TJB;DUTTON, SL
通讯作者：
DUTTON, SL
DOI：
10.1080/15287399209531623
发表时间：
1992-05-01
期刊：
JOURNAL OF TOXICOLOGY AND ENVIRONMENTAL HEALTH
影响因子：
--
作者：
GREGOTTI, C;DINUCCI, A;FAUSTMAN, EM
通讯作者：
FAUSTMAN, EM