Ultra-high Throughput, High Purity Stem Cell Isolation

超高通量、高纯度干细胞分离

• 批准号: 6647535
• 负责人: MANISH DESHPANDE
• 金额: $ 10万
• 依托单位: TERAGENICS, INC.
• 依托单位国家: 美国
• 财政年份: 2003
• 资助国家: 美国
• 起止时间: 2003-06-15 至 2003-11-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6647535
• 关键词: CD34 molecule; bioengineering /biomedical engineering; cell sorting; consumable /disposable biomedical equipment; data collection methodology /evaluation; fluid flow; hematopoietic stem cells; high throughput technology; optics; stem cells; technology /technique development; tissue /cell preparation

DESCRIPTION (provided by applicant): The goal of this project is to develop a very high-speed (1 billion cells/hour) cell sorting system with hierarchical architecture using multiple markers to achieve ultra high purity (99.999 percent) Hematopetic Stem Cells (HSC) samples derived from adult peripheral blood or bone marrow and their precursor Mesenchymal Stem Cell (MSC) samples derived from adult bone marrow. HSC/MSC derived from adult sources circumvent the ethical issues surrounding the use of embryonic stem cells while holding the promise for cell replacement therapies for diseases such as diabetes, Parkinson's Disease, spinal chord injury, and many others. Specific aims of Phase 1 of this project are: 1. Develop overall architecture of the high-speed cell sorter including the sub-systems for optics, data acquisition and control. 2. Demonstrate the feasibility of single channel switching speed of 2,000 cells/second. 3. Develop the design of micro-fabricated parallel channel array-chip, Gigasort, to achieve sorting speed of 1 billion cells/hour. 4. Develop a plan for test and validation of Teragenics' high-speed cell sorter including preliminary experimental protocols for use with specific cell markers. The core of the proposed cell sorter comprises of a microfluidic chip, Gigasort (Teragenics patent pending), which takes advantage of a rapid switching methodology coupled with a high degree of parallelism to enable the cell sorting at high speeds and a hierarchical approach to sorting to enable high sorting purity. The Gigasort chips are manufacturable using high volume microfabrication techniques and are low cost and disposable. The Gigasort chip interfaces with instrument sub-systems for optical detection and readout, control electronics, reagent storage etc and works with multiple positive and negative markers for HSC/MSC such as CD34, CD34+, Sca+, Lin-in a single hierarchical sort. The proposed project will deliver the blue print for the high speed cell sorter that delivers 3 orders of magnitude purer HSC/MSC samples 4 times faster than available methods with a cost model that allows the Gigasort microfluidic chip to be disposable, thus maintaining a fully sterile environment for clinical applications.

描述（由申请人提供）：本项目的目标是开发一种非常高速（10亿个细胞/小时）的细胞分选系统，该系统具有分层结构，使用多种标记物获得超高纯度（99.999%）的造血干细胞（HSC）样本，这些样本来自成人外周血或骨髓，以及来自成人骨髓的前体间充质干细胞（MSC）样本。源自成人来源的HSC/MSC规避了围绕使用胚胎干细胞的伦理问题，同时保持了用于诸如糖尿病、帕金森病、脊髓损伤等疾病的细胞替代疗法的希望。该项目第一阶段的具体目标是：1。开发高速细胞分选机的整体架构，包括光学、数据采集和控制子系统。2.演示单通道切换速度达到2,000个信元/秒的可行性。3.开发微加工并行通道阵列芯片Gigasort的设计，以实现10亿个细胞/小时的分选速度。4.制定Teragenics高速细胞分选仪的测试和验证计划，包括用于特定细胞标记物的初步实验方案。所提出的细胞分选仪的核心包括微流体芯片Gigasort（Teragenics专利申请中），其利用与高度并行性相结合的快速切换方法来实现高速的细胞分选，以及分级分选方法来实现高分选纯度。Gigasort芯片可使用大批量微制造技术制造，成本低，一次性使用。Gigasort芯片与用于光学检测和读出、控制电子设备、试剂存储等的仪器子系统连接，并在单个分级分类中与HSC/MSC的多个阳性和阴性标记物（例如CD 34、CD 34+、Sca+、Lin-）一起工作。拟议的项目将提供高速细胞分选仪的蓝图，该分选仪提供3个数量级的纯HSC/MSC样本，比现有方法快4倍，其成本模型允许Gigasort微流控芯片是一次性的，从而为临床应用保持完全无菌的环境。