| Summary: | The encapsulation of stem cells into alginate microspheres is an important aspect of tissue engineering or bioprinting which ensures cell growth and development. We previously demonstrated the encapsulation of stem cells using the hanging drop method. However, this conventional process takes a relatively long time and only produces a small-volume droplet. Here, an experimental approach for alginate emulsification in multistage microfluidics is reported. By using the microfluidic method, the emulsification of alginate in oil can be manipulated by tuning the flow rate for both phases. Two-step droplet emulsification is conducted in a series of polycarbonate and polydimethylsiloxane microfluidic chips. Multistage emulsification of alginate for stem cell encapsulation has been successfully reported in this study under certain flow rates. Fundamental non-dimensional numbers such as Reynolds and capillary are used to evaluate the effect of flow rate on the emulsification process. Reynolds numbers of around 0.5–2.5 for alginate/water and 0.05–0.2 for oil phases were generated in the current study. The capillary number had a maximum value of 0.018 to ensure the formation of plug flow. By using the multistage emulsification system, the flow rates of each process can be tuned independently, offering a wider range of droplet sizes that can be produced. A final droplet size of 500–1000 µm can be produced using flow rates of 0.1–0.5 mL/h and 0.7–2.4 mL/h for the first stage and second stage, respectively. © 2023 by the authors.
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