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Microfluidic device capable of medium recirculation for non-adherent cell culture
Angela R Dixon1, Shrinidhi Rajan1, Chuan-Hsien Kuo2
1Department of Biomedical Engineering, College of Engineering, University of Michigan, Ann Arbor, Michigan 48109, USA.
Biomicrofluidics
|April 23, 2014
Summary
This study introduces a novel microfluidic device for culturing non-adherent cells, enabling controlled medium recirculation and refreshing. The RECIR-REFRESH device optimizes cell differentiation by managing fluid flow and autocrine factor concentration.
Area of Science:
- Biotechnology
- Cell Biology
- Microfluidics
Background:
- Non-adherent mammalian cells require specialized culture conditions for maintenance and study.
- Existing methods for non-adherent cell culture often lack precise control over medium exchange and factor retention.
- Understanding cellular responses to controlled microenvironments is crucial for advancing stem cell research and drug discovery.
Purpose of the Study:
- To develop and validate a novel microfluidic device, RECIR-REFRESH, for the culture of non-adherent mammalian cells.
- To investigate the impact of fluid flow dynamics, specifically recirculation and refresh modes, on cell differentiation.
- To analyze the role of microfluidic device geometry in retaining autocrine factors crucial for cell signaling.
Main Methods:
- Fabrication of a microfluidic device using Braille perfusion for controlled peristaltic fluid flow.
- Utilization of computational fluid dynamics (CFD) simulations to profile flow patterns and concentration gradients.
- Culture and differentiation induction of human promyelocytic leukemia (HL-60) cells using dimethyl sulfoxide (DMSO).
- Quantification of cell surface marker CD11b and cytokine tumor necrosis factor-alpha (TNF-α) expression.
Main Results:
- The RECIR-REFRESH device successfully enabled recirculation and refreshing of medium for non-adherent cell culture.
- CFD simulations highlighted the importance of the cell culture well geometry in entrapping and retaining soluble factors.
- HL-60 cell differentiation, indicated by CD11b expression, was sensitive to perfusion rates, with a 1:1 recirculation to refresh ratio showing the highest increase.
- TNF-α levels did not show significant build-up by day 4, suggesting controlled cytokine dynamics within the device.
Conclusions:
- The RECIR-REFRESH microfluidic device provides a platform for programmable control over non-adherent cell differentiation.
- The device facilitates efficient maintenance and culture of non-adherent cells, including potential applications for hematopoietic stem cells.
- Microfluidic design parameters significantly influence cellular responses and factor retention, offering new avenues for optimizing cell culture systems.
