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

• 批准号: 8901247
• 负责人: Tony Jun Huang
• 金额: $ 28.48万
• 依托单位: PENNSYLVANIA STATE UNIVERSITY, THE
• 依托单位国家: 美国
• 财政年份: 2014
• 资助国家: 美国
• 起止时间: 2014-08-01 至 2016-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8901247
• 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; Health; Hela Cells; Hepatocyte; Human; Individual; Malaria; 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; personalized medicine; pressure; research study; single cell analysis; sperm cell; tool; trait; transcriptome sequencing; transcriptomics; voltage

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：进行高通量、高纯度、多参数细胞分选的能力对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，荧光激活的细胞分选器已经成为该领域的“金标准”技术。然而，当前的细胞分选器在处理过程中无法保持细胞的完整性 细胞分选过程。据报道，传统的细胞分选过程会显著降低许多脆弱或敏感细胞(如神经元)的细胞存活率和功能(减少30%-70%)， 干细胞、肝细胞、树突状细胞、精子细胞，甚至来自健康个体的中性粒细胞。此外，我们最近的初步结果表明，在细胞分选过程中，基因表达可以显著改变，即使是对于健壮的细胞(如HeLa细胞)。这些缺陷大大限制了细胞分选机在许多生物医学研究和临床应用中的用途，并产生了许多未得到满足的需求。例如，人类诱导的多能干细胞(IPS)为直接使用人类细胞建模人类疾病开辟了一个新的领域。它们对于药物筛选和个性化药物治疗非常有用。然而，今天仍然不可能使用细胞分选器或任何其他现有的方法以高通量、高纯度和高细胞完整性的方式分离未分化的iPS细胞。这种未得到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站立表面声波(SSAW)的高单元完整性分选器来解决这些未得到满足的需求。与传统的分选机相比，建议的SSAW细胞分选机体积更小，成本更低，预计将显著改善分选后脆弱和健壮细胞的细胞存活率、功能和基因表达。具体地说，我们将(1)开发基于SSAW的流式细胞仪，实现无鞘、多色、高通量的单细胞分析；(2)展示使用聚焦叉指换能器(f-IDT)的高通量单细胞偏转单元；(3)建立完全集成的、基于SSAW的细胞分选机系统，其性能优于最先进的细胞分选机；以及(4)展示在保持细胞完整性的情况下对诱导多能干细胞(IPS)进行分选。我们提出的基于SSAW的细胞分选机具有前所未有的能力来保持细胞的完整性，即使是脆弱的细胞，它不仅将成为现有细胞分选机的更紧凑、更实惠、更易于维护的替代品，而且还将满足基础生物医学研究以及临床诊断和治疗方面的许多未得到满足的需求。