High Precision Optical One-to-One Cell Fusion

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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.