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Published on: March 21, 2014
Effect of flow and surface conditions on human lymphocyte isolation using microfluidic chambers
Shashi K Murthy1, Aaron Sin, Ronald G Tompkins
1Center for Engineering in Medicine and Surgical Services, Massachusetts General Hospital, Harvard Medical School, Boston, Massachusetts 02114, USA.
Insights
This study developed a microfluidic device for isolating pure lymphocyte subpopulations, T and B cells, by optimizing flow conditions and surface chemistry for improved cell adhesion and reproducibility.
Area of Science:
- Immunology
- Biotechnology
- Microfluidics
Background:
- Phenotypically pure lymphocyte subpopulations are crucial for understanding immune responses.
- Isolating these cells presents challenges, especially with minimal preprocessing.
- Current methods often require extensive cell tagging, complicating analysis.
Purpose of the Study:
- To investigate microfluidic separation of T and B lymphocytes.
- To optimize flow conditions and surface chemistry for cell adhesion.
- To develop a method for isolating pure lymphocyte subpopulations with minimal preprocessing.
Main Methods:
- Utilizing microfluidic chambers coated with antibodies for cell separation.
- Examining the effect of shear stress on lymphocyte adhesion.
- Incorporating poly(ethylene glycol) chains with antibodies on chamber surfaces.
Main Results:
- Lymphocyte adhesion decreased with increasing shear stress.
- Poly(ethylene glycol) incorporation enhanced adhesion reproducibility.
- The microfluidic technique effectively isolated pure lymphocyte subpopulations from mixtures.
Conclusions:
- Microfluidic devices with optimized surface chemistry offer an effective method for lymphocyte isolation.
- This approach minimizes preprocessing, enabling purer cell subpopulations.
- The technique is valuable for studying immune responses in various conditions.
Abstract:
Phenotypically pure subpopulations of lymphocytes can provide valuable insights into the immune response to injury and disease. The isolation of these subpopulations presents unique challenges, particularly when preprocessing incubation to attach fluorescent or antibody tags is to be minimized. This paper examines the separation of T and B lymphocytes from mixtures using microfluidic chambers coated with antibodies, focusing on flow conditions and surface chemistry. The adhesion of both cell types decreases as shear stress increases irrespective of the surface chemistry. The incorporation of poly(ethylene glycol) chains along with the antibodies on the chamber surface is shown to significantly improve the reproducibility of cell adhesion and is thus an important part of the overall system design. Furthermore, this technique is shown to be an effective way of isolating highly pure subpopulations of lymphocytes from model mixtures, even when the target cell concentration is low.
