Standing Surface Acoustic Wave Based Cell Sorters for Maintaining Cell Integrity

用于保持细胞完整性的基于表面声波的细胞分选仪

基本信息

批准号：
8761977
负责人：
Tony Jun Huang
金额：
$ 28.01万
依托单位：
PENNSYLVANIA STATE UNIVERSITY, THE
依托单位国家：
美国
项目类别：
财政年份：
2014
资助国家：
美国
起止时间：
2014-08-01 至 2018-07-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/8761977
关键词：
Acoustics Address Apoptosis Biological Biomedical Research Blood Cells Blood specimen Cell Separation Cell Survival Cell Therapy Cell physiology Cells Clinical Collection Color Cytometry Data Analyses Dendritic Cells Detection Diagnostic Event Gene Expression Gene Expression Regulation Genes Genomics Gold Hela Cells Hepatocyte Human Individual Malaria Medicine Methods Modeling Nature Neurons Normal Cell Performance Phase Physiology Plasmodium Preclinical Drug Evaluation Process Production Property Proteomics Reporting Research Research Personnel Resolution Sampling Series Signal Pathway Sorting - Cell Movement Stem Cell Research Stem cells Surface Survival Rate System Techniques Technology Testing Therapeutic Tissues Transducers Tube Undifferentiated base clinical Diagnosis clinical application computerized data processing cytokine data acquisition design electric field electronic data experience fluorescence activated cell sorter device human disease improved induced pluripotent stem cell interdisciplinary collaboration metabolomics neutrophil optical fiber pressure public health relevance research study single cell analysis sperm cell tool trait transcriptome sequencing transcriptomics voltage

项目摘要

DESCRIPTION: The ability to perform high-throughput, high-purity, multi-parametric cell sorting is extremely important for many biomedical studies and clinical applications. In the past few decades, fluorescence-activated cell sorters have become the "gold standard" technique in the field. However, current cell sorters suffer from an inability to maintain cell integrity during the cell- sorting process. Conventional cell-sorting processes are reported to significantly reduce cell viability and function (30-70% reduction) for many fragile or sensitive cells such as neurons, stem cells, liver cells, dendritic cells, sperm cells, and even neutrophils from healthy individual. In addition, our recent preliminary results indicate that gene expression can be significantly altered during the cell-sorting process, even for robust cells (such as HeLa cells). These drawbacks significantly limit the usefulness of cell sorters in many biomedical studies and clinical applications and have created many unmet needs. For example, human induced pluripotent stem (iPS) cells have opened a new field for modeling human diseases using human cells directly. They can be extremely useful for drug screening and personalized medicine. However, today it is still impossible to use cell sorters or any other existing methods to isolate undifferentiated iPS cells in a high-throughput, high-purity, and high-cell-integrity manner. This unmet need has significantly hindered progress in stem cell research and therapy. Our objective is to address these unmet needs by demonstrating standing surface acoustic wave (SSAW) based, high-cell-integrity sorters. When compared to conventional sorters, the proposed SSAW cell sorter is substantially smaller and less expensive, and is expected to significantly improve post-sorting cell viability, function, and gene expression for both fragile and robust cells. In particular, we will (1) develop a SSAW-based flow cytometer that achieves sheathless, multi-color, high-throughput single-cell analysis; (2) demonstrate a high-throughput, single-cell deflecting unit using focused interdigital transducers (f-IDTs); (3) establish a fully integrated, SSAW-based cell sorter system proven with human blood samples to outperform a state-of-the-art cell sorter; and (4) demonstrate sorting of induced pluripotent stem (iPS) cells with maintained cell integrity. With unprecedented capabilities to maintain cell integrity, even for fragile cells, our proposed SSAW-based cell sorter will not only become a more compact, affordable, and easy-to-maintain replacement to the existing cell sorters, but also fill many unmet needs in both fundamental biomedical research and clinical diagnosis and therapeutics.

描述：执行高通量、高纯度、多参数细胞分选的能力对于许多生物医学研究和临床应用极其重要。在过去的几十年里，荧光激活细胞分选仪已成为该领域的“金标准”技术。然而，当前的细胞分选仪无法在分选过程中保持细胞完整性。细胞分选过程。据报道，传统的细胞分选过程会显着降低许多脆弱或敏感细胞（例如神经元）的细胞活力和功能（减少 30-70%）。来自健康个体的干细胞、肝细胞、树突状细胞、精子细胞，甚至中性粒细胞。此外，我们最近的初步结果表明，即使对于稳健的细胞（例如 HeLa 细胞），基因表达在细胞分选过程中也可能发生显着改变。这些缺点极大地限制了细胞分选仪在许多生物医学研究和临床应用中的实用性，并产生了许多未满足的需求。例如，人类诱导多能干（iPS）细胞开辟了直接使用人类细胞模拟人类疾病的新领域。它们对于药物筛选和个性化医疗非常有用。然而，今天仍然不可能使用细胞分选仪或任何其他现有方法以高通量、高纯度和高细胞完整性的方式分离未分化的iPS细胞。这种未满足的需求严重阻碍了干细胞研究和治疗的进展。我们的目标是通过展示基于驻表面声波 (SSAW) 的高细胞完整性分选机来解决这些未满足的需求。与传统分选仪相比，所提出的 SSAW 细胞分选仪体积更小、成本更低，并且有望显着改善脆弱细胞和健壮细胞的分选后细胞活力、功能和基因表达。具体来说，我们将（1）开发基于SSAW的流式细胞仪，实现无鞘、多色、高通量单细胞分析； (2) 展示使用聚焦叉指换能器 (f-IDT) 的高通量单细胞偏转装置； (3) 建立一个完全集成的、基于 SSAW 的细胞分选系统，经人体血液样本证明其性能优于最先进的细胞分选系统； (4) 展示诱导多能干 (iPS) 细胞的分选并保持细胞完整性。我们提出的基于 SSAW 的细胞分选仪具有前所未有的保持细胞完整性的能力，即使对于脆弱的细胞也是如此，它不仅将成为现有细胞分选仪的更紧凑、价格实惠且易于维护的替代品，而且还可以满足许多未满足的需求。基础生物医学研究和临床诊断和治疗。