A human-derived prostate co-culture microtissue model using epithelial (RWPE-1) and stromal (WPMY-1) cell lines.
A human-derived prostate co-culture microtissue model using epithelial (RWPE-1) and stromal (WPMY-1) cell lines.
复制标题
使用上皮细胞 (RWPE-1) 和基质细胞 (WPMY-1) 细胞系的人源性前列腺共培养微组织模型。
DOI：
10.1016/j.tiv.2019.05.023
复制
发表时间：
2019
期刊：
影响因子：
--
通讯作者：
Boekelheide,Kim
中科院分区：
文献类型：
--
作者：
Dent,MatthewP;Madnick,SamanthaJ;Hall,Susan;VantangoliPolicelli,Marguerite;Bars,Chloe;Li,Hui;Amin,Ali;Carmichael,PaulL;Martin,FrancisL;Boekelheide,Kim
前列腺组织的发育和正常功能取决于间质和上皮间室之间的信号相互作用。由这两种成分组成的前列腺微组织的发展可以帮助识别可能对人类造成不良影响的物质暴露，作为非动物风险评估的一部分。本研究利用无支架水凝胶培养由人源性基质（WPMY-1）和上皮（RWPE-1）细胞系组成的前列腺微组织，并利用免疫组织化学、光镜和qRT-PCR对其进行了表征。在播种后5 天内，微组织自组织成球体，由基质WPMY-1细胞核心组成，周围是上皮RWPE-1细胞。RWPE-1层反映中间前列腺上皮，表达腔上皮细胞（PSA高表达）和基底上皮细胞（细胞角蛋白5/6和14高表达）的特征。显微组织对雄激素（二氢睾酮，DHT）和抗雄激素（氟他胺）的反应也进行了研究。用二氢睾酮、氟他胺或二氢睾酮和氟他胺的混合物治疗表明，微组织的形态和自组织依赖于雄激素。qRT-PCR数据显示，饱和浓度的DHT增加了雌激素受体（ESR1和ESR2）编码基因的表达，降低了CYP1B1的表达，但不影响雄激素受体的表达。随着进一步的开发和优化，RWPE-1/WPMY-1微组织可在非动物风险评估中发挥重要作用。
The development and normal function of prostate tissue depends on signalling interactions between stromal and epithelial compartments. Development of a prostate microtissue composed of these two components can help identify substance exposures that could cause adverse effects in humans as part of a non-animal risk assessment. In this study, prostate microtissues composed of human derived stromal (WPMY-1) and epithelial (RWPE-1) cell lines grown in scaffold-free hydrogels were developed and characterized using immunohistochemistry, light microscopy, and qRT-PCR. Within 5 days after seeding, the microtissues self-organized into spheroids consisting of a core of stromal WPMY-1 cells surrounded by epithelial RWPE-1 cells. The RWPE-1 layer is reflective of intermediate prostatic epithelium, expressing both characteristics of the luminal (high expression of PSA) and basal (high expression of cytokeratins 5/6 and 14) epithelial cells. The response of the microtissues to an androgen (dihydrotestosterone, DHT) and an anti-androgen (flutamide) was also investigated. Treatment with DHT, flutamide or a mixture of DHT and flutamide indicated that the morphology and self-organization of the microtissues is androgen dependent. qRT-PCR data showed that a saturating concentration of DHT increased the expression of genes coding for the estrogen receptors (ESR1 and ESR2) and decreased the expression of CYP1B1 without affecting the expression of the androgen receptor. With further development and optimization RWPE-1/WPMY-1 microtissues can play an important role in non-animal risk assessments.