Rare Cell Isolation for Morphological & Molecular Study

描述(申请人提供)：阐明与癌症发展相关的遗传变化和分子图谱的第一步是从数百万正常细胞的背景中分离罕见的肿瘤细胞，或将不同种类的细胞群体划分为亚群。为此，我们提出了一个通用的、基于介电泳法的细胞分离和分离平台，该平台是NCI IMAT资助的研究的产物，目前正在M.D.安德森癌症中心(MDACC)进行。该项目的目标是开发一种能够分离人类细胞并以熟悉的幻灯片格式呈现它们的设备，既适合细胞形态分析，这仍然是癌症诊断和分期的黄金标准，也可以作为新兴分子分析方法的前端。 所提出的电涂片设备利用专有的介电泳电极阵列配置和频率方案来悬浮细胞。这种阵列被配置为使不同类型的细胞以空间上不同的特征带附着在载玻片上，使该设备成为分离和分组稀有细胞的理想设备。根据MDACC的独家许可，ADEPTAS Inc.将设计适当的电极和控制电子设备，设计样品室以允许可选的磁电性分离组件，组装设备，并使用测试颗粒执行初步概念验证测试。 该装置的生物相关性测试将在MDACC进行。MDACC的实验将利用各种培养的肿瘤细胞系、全血和黄褐色涂层样本，以证明电涂片根据细胞的介电泳性将细胞亚群分离并捕获到特征条带的能力。关键实验将包括从添加了添加剂的血细胞样本中分离肿瘤细胞，以测试电子涂片法的检测极限。MDACC小组还将测试电涂片与传统细胞染色方法的兼容性、病理学家进行的细胞形态评估和最新的自动玻片计数细胞术方法，以及包括MALDI-TOF MS在内的分子分析方法。这一第一阶段SBIR项目的成功完成将为该设备的第二阶段应用和商业化奠定基础，该设备应立即适用于生命科学领域的病理学家和研究人员。

DESCRIPTION (provided by applicant): A first step in the elucidation of the genetic changes and molecular profiles associated with the development of cancer is the isolation of rare tumor cells from a background of millions of normal cells, or the fractionation of a heterogeneous cell population into subpopulations. To this end we propose a versatile, dielectrophoresis-based cell fractionation and isolation platform that is an outgrowth of NCI IMAT funded studies currently underway at M.D. Anderson Cancer Center (MDACC). The goal of the project is to develop a device capable of separating human cells and presenting them in a familiar slide format appropriate for both cytomorphological analysis, which is still the gold standard for cancer diagnosis and staging, as well as a format that can serve as a front-end for emerging molecular analysis methods. The proposed electrosmear device utilizes a proprietary dielectrophoretic electrode array configuration and frequency scheme to levitate cells. This array is configured so that different cell types adhere to the slide in spatially distinct, characteristic bands, making the device ideal for isolating and grouping rare cells. Under an exclusive license from MDACC, Adeptas Inc. will design the appropriate electrodes and controlling electronics, design the sample chamber to permit an optional magnetophoretic separation component, assemble the device, and perform initial proof of concept tests with test particles. The biologically relevant testing of the device will be conducted at MDACC. Experiments at MDACC will utilize a variety of cultured tumor cell lines, whole blood, and buffy coat samples to demonstrate the ability of the electrosmear to isolate and trap cell subpopulations into characteristic bands based upon their dielectrophoretic properties. Key experiments will involve the isolation of tumor cells from spiked blood cell samples to test the limit of detection of the electrosmear method. The MDACC group will also test electrosmear slide compatibility with traditional cell staining methods, cytomorphological evaluation by both a pathologist and more recent automated slide counting cytometry methods, and molecular analysis methods including MALDI-TOF MS. Successful completion of this Phase I SBIR project should provide a basis for a Phase II application and commercialization of the device, which should be immediately applicable by pathologists and researchers in the life sciences.