IN VITRO LOW-SHEAR THREE-DIMENSIONAL BLADDER TUMOR MODEL

研制了一种新型装置--大展宽旋转壁容器(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生产的三个 低流体剪切力下的三维细胞聚集体(球体) 条件，模拟微重力的一个方面。血管使细胞旋转 和在围绕水平轴线的盘状室中的培养介质 没有顶层空间。在膜上保持足够的气体交换 排列在腔室的一面。在与美国宇航局的合作下， 细胞诊断公司(CDI)研究了人类肿瘤细胞的生长 Harv中的三维球体线条(胶质瘤和前列腺线) 仪器。利用分子生物标记物，CDI及其合作者发现了DNA 独特的肿瘤生物标记物的含量、F-肌动蛋白组织和表达 与体内癌症分子变化相关，但与单层培养无关 从固定培养的组织培养瓶中提取。这项提议旨在 在膀胱细胞系肿瘤模型中证实和推广我们的观察结果 翼专利肿瘤标志物；粘蛋白，F/G-肌动蛋白组织， 癌基因、肿瘤抑制基因、表皮生长因子和 用于评估已建立的已知细胞系的比较的受体探针 在Matrigel(TM)细胞生长的基础上生长为二维单层 使用HARV生产的衬底和3-D液体生长的球体 技术要使用的细胞系包括T-24、J-82和几个 我们的合作者雷兹尼科夫博士在 威斯康星大学，已经被广泛地描述了 从细胞遗传学上讲。其目标是研究体外培养的膀胱细胞 在时间上对原位表达基因的差异、定量 和表型生物标记物。目标将是建立生物标记物档案 保真度已知的体内膀胱癌结果建立在 文学。该模型最终将应用于增长和扩张。 用于新治疗药物定制评估的小肿瘤活组织检查。 正在开发3D椭球体技术将在 体外模型用于评估新的治疗方法和研究新的作用机制 以及治疗癌症的新疗法。

A 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 tumor cell lines (Glioma and Prostate) as three dimensional spheroids in the HARV apparatus. Using molecular biomarkers, CDI and its collaborators found DNA content, F-actin organization, and expression of unique tumor biomarkers correlated to in vivo cancer molecular changes, but not monolayer cultures from stationary grown tissue culture flasks. This proposal is designed to confirm and extend our observations in a bladder cell line tumor model wing proprietary tumor markers; mucins, F/G-actins organization, oncogenes, tumor suppressor genes, and epidermal growth factor and receptor probes to assess the comparisons of established known cell lines grown as two dimensional monolayers grown on Matrigel(TM) cell growth substrate versus 3-D liquid grown spheroids produced using the HARV technology. The cell lines to be employed include T-24, J-82, and several unique cell lines developed by our collaborator, Dr. Reznikoff at the University of Wisconsin, which have been extensively characterized cytogenetically. The goal is to study in vitro grown bladder cells temporally for differential, quantitatiion of in situ expressed genetic and phenotypic biomarkers. A goal will be to establish biomarker profile fidelity to known in vivo bladder tumor results established in the literature. The model will ultimately be applied to growth and expansion of small tumor biopsies for custom evaluation of new therapeutic agents. The 3D spheroid technology being developed will be provide a useful in vitro model to assess new treatments and study mechanisms of action of new and novel treatments for cancer.