IN VITRO LOW-SHEAR THREE-DIMENSIONAL BLADDER TUMOR MODEL

体外低剪切三维膀胱肿瘤模型

• 批准号: 3493484
• 负责人: ROBERT W VELTRI
• 金额: $ 4.99万
• 依托单位: UROCOR, INC.
• 依托单位国家: 美国
• 财政年份: 1993
• 资助国家: 美国
• 起止时间: 1993-09-15 至 1994-03-14
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/3493484
• 关键词: biological models; biomarker; bladder neoplasm; cell aggregation; cell growth regulation; clone cells; gravity; mass tissue /cell culture

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.

本文介绍了一种新型的高宽比旋转壁容器（High Aspect Rotating Wall Vessel，HARV） 由美国宇航局在约翰逊航天中心，休斯敦，得克萨斯州，产生三个 低流体剪切下的三维细胞聚集体（球状体） 模拟微重力的一个方面。血管旋转细胞 和培养基在盘形室中围绕水平轴， 零顶空 通过膜维持充分的气体交换 在房间的一面。在与NASA的合作下， CytoDiagnostics，Inc. (CDI)研究了人类肿瘤细胞的生长 线（胶质瘤和前列腺）作为三维球体在HARV 设备.利用分子生物标志物，CDI及其合作者发现了DNA 独特肿瘤生物标志物的含量、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 从静止生长的组织培养瓶中。这项建议旨在 证实并扩展了我们在膀胱细胞系肿瘤模型中的观察结果 翼专有肿瘤标志物;粘蛋白，F/G-肌动蛋白组织， 癌基因、肿瘤抑制基因和表皮生长因子， 受体探针，以评估已建立的已知细胞系的比较 在Matrigel（TM）细胞生长上生长为二维单层 基质与使用HARV产生的3-D液体生长球状体 技术.待使用的细胞系包括T-24、J-82和几种细胞系。 由我们的合作者Reznikoff博士开发的独特细胞系， 威斯康星州的大学，这已经广泛的特点 细胞遗传学目的是研究体外培养的膀胱细胞 原位表达基因的时间形成差异、定量 和表型生物标志物。目标是建立生物标志物谱 与已知的体内膀胱肿瘤结果的保真度， 文学该模型最终将应用于增长和扩张 小肿瘤活检用于新治疗药物的定制评估。 正在开发的3D球体技术将提供一个有用的， 体外模型，以评估新的治疗方法和研究新的 以及癌症的新疗法。