Abstract
Cancer cell–immune cell hybrids and cancer immunotherapy have attracted much attention in recent years. The design of efficient cell pairing and fusion chips for hybridoma generation has been, subsequently, a subject of great interest. Here, we report a three-layered integrated Microfluidic Flip-Chip (MFC) consisting of a thin through-hole membrane sandwiched between a mirrored array of microfluidic channels and saw-tooth shaped titanium electrodes on the glass. We discuss the design and operation of MFC and show its applicability for cell fusion. The proposed device combines passive hydrodynamic phenomenon and gravitational sedimentation, which allows the transportation and trapping of homotypic and heterotypic cells in large numbers with pairing efficiencies of 75~78% and fusion efficiencies of 73%. Additionally, we also report properties of fused cells from cell biology perspectives, including combined fluorescence-labeled intracellular materials from THP1 and A549, mixed cell morphology, and cell viability. The MFC can be tuned for pairing and fusion of cells with a similar protocol for different cell types. The MFC can be easily disconnected from the test setup for further analysis.
Highlights
The Microfluidic Flip-Chip (MFC) uses a two-step protocol for the cell loading process
The presence of a thin membrane might look like an issue concerning leakage between the two trapping channels, a thorough experimental verification has been carried out, and it was observed that at optimum flow rates, as discussed in the previous sections, there is a negligible cross channel flow observed at lower flow rates ensuring that the cell trapping process is unaffected
Microfluidics has enabled the precise manipulation of cells
Summary
Cellular vaccines were produced and described a decade ago, based on the cell fusion of dendritic cells (DCs) and cancer cells to offer hybrid cells sharing a united cytoplasm but keeping the identity of dual nuclei [2,3,4]. Cellular fusion is a process in which two or more cells are merged in an asexual way producing a hybrid cell. Extending the use of such hybridomas has led to the development of a tool called cancer immunotherapy [5]. In addition to cancer immunotherapy [6], applications of cell fusion have long been discussed extensively in a variety of fields such as hybridoma generation [7,8], reprogramming of somatic cells [9], and mammal cloning [10,11]. Cell fusion can be categorized primarily into three types, viz, poly-ethylene glycol (PEG) based fusion (chemical fusion) [12], fusion via viruses (biological) [13], and electroporation (physical) [14]
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