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Endothelialized Microfluidics for Studying Microvascular Interactions in Hematologic Diseases
Published on: June 22, 2012
Detection of circulating endothelial cells via a microfluidic disk
Ken-Chao Chen1, Tai-Ping Lee, Yu-Cheng Pan
1Institute of Applied Mechanics, National Taiwan University Hospital, College of Medicine, National Taiwan University, Taipei, Taiwan.
Insights
Detecting rare circulating endothelial cells (CECs) is challenging. This study introduces an economical microfluidic disk for CEC detection, showing higher counts in systemic lupus erythematosus patients, indicating its clinical viability.
Area of Science:
- Biomedical Engineering
- Cell Biology
- Clinical Diagnostics
Background:
- Circulating endothelial cells (CECs) are rare blood markers linked to diseases.
- Detecting CECs amidst peripheral blood mononuclear cells (PBMCs) presents significant challenges due to low ratios.
- A novel microfluidic disk system is developed for efficient CEC isolation and detection.
Purpose of the Study:
- To develop an economical microfluidic disk for detecting circulating endothelial cells (CECs).
- To validate the system's efficacy using a model cell system and clinical samples.
- To assess the diagnostic potential of CEC counts in systemic lupus erythematosus (SLE).
Main Methods:
- Utilized immunomagnetic beads and a custom magnet on a microfluidic disk for cell enrichment.
- Employed specific antibodies (anti-CD146-phycoerythrin for CECs, anti-CD45-FITC for PBMCs) for cell labeling.
- Validated detection efficiency with model cells (HUVECs in PBMCs) and compared results with flow cytometry.
Main Results:
- Achieved 95% detection of spiked human umbilical vein endothelial cells (HUVECs) in PBMCs.
- Demonstrated high yield even at low cell ratios (60 HUVECs in <10^4 PBMCs).
- Found significantly higher CEC counts in SLE patients (61.1) compared to healthy donors (31.2).
Conclusions:
- CEC count serves as a suitable biomarker for systemic lupus erythematosus symptoms.
- The developed microfluidic disk system offers a viable platform for CEC detection.
- This technology has potential for broader clinical diagnostic applications.
Background:
Circulating endothelial cells (CECs) in the blood are rare but have been shown to be associated with various diseases. With the ratio of CECs to peripheral blood mononuclear cells (PBMCs) less than 1 part per thousand, their separation from PBMCs and detection are challenging. We present a means of detecting CECs from PBMCs via an economical microfluidic disk with a model cell system [human umbilical vein endothelial cells (HUVECs) in PBMCs], along with demonstration of its efficacy clinically.
Methods:
To enrich these rare cells, we used immunomagnetic beads and a tailor-made magnet on the disk. CEC-simulating HUVECs, as target cells, were stained with primary anti-CD146-phycoerythrin antibody and bound with secondary antibody on antiphycoerythrin magnetic beads. PBMCs served as nontarget cells and were labeled with anti-CD45-FITC antibody.
Results:
When hundreds of HUVECs were mixed in 10(6) PBMCs, 95% of spiked HUVECs were detected. This yield also held for 60 HUVEC in <10(4) PBMCs. We compared data from flow cytometry with that from the disk: CEC counts in 50 μL blood from patients with systemic lupus erythematosus were 61.1 (21.5), significantly higher (P < 0.01) than those of healthy donors, 31.2 (13.3).
Conclusions:
The count of CECs is a suitable marker for symptoms of systemic lupus erythematosus. The microfluidic disk system should be a viable platform for detection of CECs.
