Preparation of mammalian cell-enclosing subsieve-sized capsules (<100 microm) in a coflowing stream

The droplet breakup technique with an immiscible liquid coflowing stream was investigated for the preparation of mammalian cell-enclosing subsieve-sized capsules of less than 100 microm in diameter. The major parts of the droplet generation device were a needle of several hundred micrometers in diameter for extruding the cell-suspending sodium alginate aqueous solution and a tubule of 2.5 mm in diameter through which the extruded alginate solution flowed into ambient immiscible liquid paraffin. The needle was positioned upstream in the vicinity of the coaxial tubule. The droplet diameter of the viscous sodium alginate aqueous solution could be controlled from several dozen to several hundred micrometers by changing the velocities of the inner and ambient fluids and the diameter of the needle. By utilizing a 300-microm diameter needle, CHO-K1 cell-enclosing droplets of 48 +/- 8 microm in diameter were obtained by extruding a cell-suspending sodium alginate solution at a velocity of 1.2 cm/sec into the ambient liquid paraffin flowing at a velocity of 23.5 cm/sec. The breakup process did not influence the viability of the enclosed cells, since more than 95% of the CHO-K1 cells remained alive after the enclosing process.