High Precision Optical One-to-One Cell Fusion

描述(申请人提供)：在过去的十年里，细胞融合已经成为分析杂交瘤抗体产生、癌症免疫治疗、基因导入、染色体定位、克隆哺乳动物和干细胞研究的强大工具。到目前为止，细胞融合技术一直基于化学和物理方法。然而，使用这些技术，某些类型的刺激造成的全细胞损伤是不可避免的，而且由于集体治疗，不可能融合特定目的所需的特定细胞。因此，Maxwell Sensors Inc.提议开发一种新型的光学细胞融合(OC-Fusion)芯片和系统，该芯片和系统结合了微滴和光学融合技术，以非常高的精度快速产生数百或数千个混合细胞。微滴方法-快速而准确地将相互接触的细胞配对在一个微小的液滴中。激光精确地融合了细胞接触处的一个点。OC-Fusion的优势在于：1.多功能液滴技术可在微滴内快速配对1：1接触的选定细胞；这一过程不依赖于任何自然的细胞接触、特定的细胞受体或劳动密集型光学捕获。2.使用图像辅助激光系统扫描液滴阵列，并在点接触处用紫外线微束精确融合两个细胞，可产生成百上千个混合细胞。为了实现拟议项目的目标，第一阶段的具体目标是设计和开发OC-Fusion芯片和系统，表征系统性能，制定方案，进行细胞-细胞电融合，以融合(1)培养的中国仓鼠卵巢(CHO-K1)细胞，(2)脾细胞和骨髓瘤。与公共健康相关：Maxwell Sensors Inc.计划开发一种紧凑型单细胞融合芯片和系统，该芯片和系统能够为各种生物技术和生物医学应用产生数百或数千个融合细胞。光学融合，即使用激光脉冲融合两个或更多活细胞的过程，是一项应用广泛的不断发展的技术。这项技术是癌症免疫治疗、单克隆抗体生成和干细胞研究不可或缺的一部分，在干细胞研究中，它让研究人员更深入地了解基因如何引导蛋白质合成。这项技术还允许研究人员创造大量不同的疾病特异性抗体，并帮助科学家克隆哺乳动物。

DESCRIPTION (provided by applicant): In the past decade, cell fusion has become a powerful tool for the analysis of hybridoma antibody production, cancer immunotherapy, gene transfection, chromosomal mapping, cloning mammals, and stem cell research. Until now, cell fusion techniques have been based on chemical and physical methods. Using these techniques, however, whole cell damage caused by some types of stimulation was unavoidable and a fusion of specific cells that are required for particular purposes was not possible because of the collective treatment. Therefore, Maxwell Sensors Inc. proposes to develop a novel Optical Cell Fusion (OC-Fusion) chip and system that combines the technologies of picodroplet and optical fusion to rapidly generate hundreds or thousands of hybrid cells with very high precision. The picodroplet method - rapidly and accurately pairs cells in contact with each other in a tiny droplet. A laser - precisely fuses a point at cell contact. The advantages of OC-Fusion are: 1. a versatile droplet technique rapidly pairs 1:1 selected cells in contact inside a picodroplet; the process does not depend on any natural cell contact, specific cell receptors, or labor-intensive optical trapping. 2. hundreds or thousands of hybrid cells can be generated with an image-assisted laser system used to scan the droplet array and precisely fuse the two cells at point contact (< 3<m) with a UV microbeam. In order to achieve the goal of the proposed project, the Phase I specific aims are to design and develop the OC-Fusion chip and system, characterize the system performance, develop protocol, perform and validate cell-cell electrofusion to fuse (1) cultured Chinese hamster ovary (CHO-k1) cells, and (2) splenocyte and myeloma. PUBLIC HEALTH RELEVANCE: Maxwell Sensors Inc. proposes to develop a compact single-cell fusing chip and system that enables to generate hundreds or thousands of fused cells for a variety of biotechnology and biomedical applications. Optical fusion, the process of using laser pulses to fuse two or more living cells, is a growing technology with a wide variety of applications. The technique is integral to cancer immunotherapy, monoclonal antibody generation and stem cell research where it affords researchers greater insight into how genes guide protein synthesis. The technique also allows researchers to create large quantities of different disease-specific antibodies and helps scientists clone mammals.