Microfluidic Cell-squeezing Technology for B Cell-based Vaccine

Rapid mechanical deformation of cells has become a promising, vector-free method for intracellular delivery of nanomaterials and macromolecules, which opens new possibilities for cellular vaccine. Creative Biolabs has developed a microfluidic cell-squeezing device that introduces specific antigens into the B cells of the immune system, providing a new approach to the development and implementation of antigen presenting cell vaccines.

Why Use Cell Squeezing as a Microfluidic Intracellular Delivery Platform?

Delivery of macromolecules into the cytoplasm is an essential step in research applications like cell therapy. However, macromolecules are difficult to penetrate into the cytoplasm. Many techniques have been developed to promote membrane disruption or endocytic delivery, such as using polymer nanoparticles, liposomes or chemical modifications. In these cases, delivery efficiency and cell viability generally depend on the structure and cell type of the target molecule. In addition, membrane perforation methods, such as electroporation and sonication, are an attractive alternative. However, they are known to cause low cell viability and may be limited by the charge of the target delivery material.

Rapid mechanical deformation of cells has become a promising, vector-free method for intracellular delivery of nanomaterials and macromolecules, which relies on mechanical disruption of the cell membrane to promote cytosolic delivery of the substances present in the surrounding buffer. This technology has been used for more than 20 cell types, including naive immune cells and embryonic stem cells.

How Does a Microfluidic Cell-squeezing Device Work?

Cell-disrupting pressure change is achieved by passing the cells through a narrow opening in the microfluidic device which consists of a channel etched into the wafer. As the cells pass through the device, the channel width gradually narrows. The flexible membrane of the cell allows it to change shape and become thinner and longer, allowing it to be squeezed and pass through. The forced rapid change in cell shape temporarily creates a hole in the membrane without damaging or killing the cell. At the same time, the target materials can enter the cell through the holes in the membrane. When the cells return to their normal shape, the holes in the membrane are closed.

Schematic of the Microfluidic Cell-squeezing Device – Creative Biolabs

Fig.1 Schematic of the Microfluidic Cell-squeezing Device.

Microfluidic Cell-squeezing Technology for B Cell-based Vaccine Production

Clinical studies of cell-based vaccines have focused on dendritic cells (DC) ("professional" APCs). However, dendritic cells have many shortcomings as a platform for clinical use, such as the relative lack of human blood, short lifespan, the inability to proliferate and complex subgroup heterogeneity, which have led to other cell types being considered for cell-based APC vaccines, including macrophages and B cells. B cells are promising candidate APCs to elicit antigen-specific T cells in vivo and in vitro. However, a significant obstacle to the use of B cells as APCs compared to "professional" APCs is their low ability to take up non-specific antigens.

Therefore, scientists of Creative Biolabs use microfluidic cell-squeezing devices to promote direct cytosolic delivery of whole proteins into live B cells through transient plasma membrane perforation. The "cell extrusion" process creates transient holes in the plasma membrane that enable the entire protein to pass from the surrounding medium cells to the B cells by mechanical manipulation.

Service Process

Microfluidic Cell-squeezing Technology for Cell-based Vaccine Production – Creative Biolabs

Creative Biolabs’ cell squeezing platform provides the possibility to prime autologous B-cells for in vitro CTL expansion and the development of B cell-based vaccines. Please feel free to contact us for more details.


  1. Sharei A; et al. Cell squeezing as a robust, microfluidic intracellular delivery platform. J Vis Exp. 2013, (81): e50980.

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