3D Printed Microfluidic Spiral Separation Device for Continuous, Pulsation-Free and Controllable CHO Cell Retention

Area of Science:

• Biotechnology
• Process Engineering
• Microfluidics

Background:

• Continuous bioprocessing requires effective cell retention for extended cultivation.
• Microfluidic systems offer precise particle manipulation for cell separation.
• Existing systems lack separation control and require pulsation-free flow, often unachievable with standard pumps.

Purpose of the Study:

• To develop and evaluate a 3D printed continuous cell separation system for bioprocesses.
• To achieve high separation efficiency and operator-controlled separation adjustment.
• To integrate pulsation dampening for stable, continuous operation with standard pumps.

Main Methods:

• Design and 3D printing of a spiral cell separation device for CHO-K1 cells.
• Optical evaluation and experimental validation with cell cultures.
• Development of integrated 3D printed buffer devices to minimize pump pulsations.
• System integration with bioreactors for continuous operation.

Main Results:

• Achieved over 95% separation efficiency for Chinese hamster ovary (CHO-K1) cells at high cell densities (up to 20 × 10^6 cells mL^-1).
• Demonstrated adjustable separation efficiency through control of inlet and outlet flow rates.
• Successfully integrated pulsation dampeners, reducing dead volume and enabling stable flow with peristaltic pumps.
• Developed a fully integrated system for continuous, pulsation-free cell retention connected to bioreactors.

Conclusions:

• The 3D printed spiral separator provides efficient and controllable cell separation for continuous bioprocessing.
• Integrated pulsation dampening is crucial for stable microfluidic cell separation with standard pumps.
• This technology enables enhanced cell retention and process optimization in bioreactors.