LOW-SHEAR 3-DIMENSIONAL IN VITRO PROSTATE TUMOR MODEL

一种新的装置--高纵横比旋转壁容器(HARV)， 由美国国家航空航天局在德克萨斯州休斯顿的约翰逊航天中心开发，生产 低流体剪切下的三维细胞聚集体(球体) 条件，模拟微重力的一个方面。船在旋转 绕水平轴的盘状室中的细胞和培养液 头顶空间为零。在整个过程中保持足够的气体交换 隔膜衬砌在腔室的一侧。在与以下公司的合作下 美国国家航空航天局细胞诊断公司(CDI)研究了人脑胶质瘤的生长 和前列腺细胞系作为三维球体在Harv 仪器。利用分子生物标记物，CDI及其合作者发现 独特肿瘤的DNA含量、肌动蛋白组织和表达 生物标记物与体内前列腺癌分子变化相关，但 不是来自固定培养的组织培养瓶的单层培养。这 建议书旨在证实和扩大我们在这方面的观察 使用专有肿瘤标记物、癌基因、肿瘤的前列腺癌模型 抑制基因和雄激素受体探针来评估比较 球体体外生长与单层和前列腺核的比较 由我们实验室处理的活组织检查。拟采用的细胞系 包括将要生长PC3-PF、PC3-MF和LNCaP前列腺细胞系 作为单分子膜和三维球体使用HARV技术 并对原位进行了时间上的差异性、定量化研究 使用CAS-200图像分析系统表达的生物标志物。曾经是一个 优化的中间生物标志物剖面，高保真 体内肿瘤建立后，将用该模型对选定的 化疗药物[苏拉明、紫杉醇、非那雄胺]以评估 该模型在评估其治疗潜力方面的应用。这个 正在开发的3D椭球体技术将在体外提供有用的 评估新的治疗方法和研究作用机制的模型 前列腺癌。

A new and novel apparatus, the High Aspect Rotating-Wall Vessel (HARV), developed by NASA at the Johnson Space Center, Houston, TX, produces three dimensional cellular aggregates (spheroids) under low fluid shear conditions, an aspect of simulated microgravity. The vessel rotates cells and culture medium in a disk-shaped chamber about a horizontal axis with zero headspace. Adequate gas exchange is maintained across a membrane lining one face of the chamber. Under a collaboration with NASA, CytoDiagnostics, Inc. (CDI) has studied the growth of human glioma and prostate cell lines as three dimensional spheroids in the HARV apparatus. Using molecular biomarkers, CDI and its collaborators found DNA content, actin organization, and expression of unique tumor biomarkers correlated to in vivo prostate cancer molecular changes, but not monolayer cultures from stationary grown tissue culture flasks. This proposal is designed to confirm and extend our observations in this prostate tumor model using proprietary tumor markers, oncogenes, tumor suppressor genes, and androgen receptor probes to assess the comparisons of the in vitro growth of spheroids versus monolayers and prostate core biopsies processed by our laboratory. The cell lines to be employed include PC3-PF, PC3-MF, and LNCaP prostate cell lines which will be grown as monolayers and three dimensional spheroids using the HARV technology and studied temporally for differential, quantitation of in situ expressed biomarkers using a CAS-200 image analysis system. Once an optimized intermediate biomarker profile with high fidelity to the in vivo tumor is established, the model will be used to assess selected chemotherapeutic agents [suramin, taxol, finasteride] to assess the utility of the model for evaluation of their therapeutic potential. The 3D spheroid technology being developed will provide a useful in vitro model to assess new treatments and study mechanisms of action for prostate cancer.