Design and validation of an endothelial progenitor cell capture chip and its application in patients with pulmonary arterial hypertension

• Published in: Journal of molecular medicine (Berlin, Germany) Vol. 89; no. 10; pp. 971 - 983
• Main Authors: Hansmann, Georg, Plouffe, Brian D., Hatch, Adam, von Gise, Alexander, Sallmon, Hannes, Zamanian, Roham T., Murthy, Shashi K.
• Format: Journal Article
• Language: English
• Published: Berlin/Heidelberg Springer-Verlag 01.10.2011; Springer; Springer Nature B.V
• Subjects: Adult; Aged; Aging - pathology; Biological and medical sciences; Biomedical and Life Sciences; Biomedicine; Body Mass Index; Case-Control Studies; Cell Count; Cell Separation; Endothelial Cells - pathology; Equipment Design; Familial Primary Pulmonary Hypertension; Female; General aspects; Human Genetics; Humans; Hypertension, Pulmonary - blood; Hypertension, Pulmonary - diagnosis; Hypertension, Pulmonary - pathology; Internal Medicine; Male; Medical sciences; Microfluidic Analytical Techniques - instrumentation; Microfluidic Analytical Techniques - methods; Middle Aged; Molecular Medicine; Original Article; Pneumology; Postmenopause - blood; Pulmonary hypertension. Acute cor pulmonale. Pulmonary embolism. Pulmonary vascular diseases; Reproducibility of Results; Stem Cells - pathology; Bioengineering; Vascular disease; Biomarker; Diagnostics; Bone marrow; Bedside test; Progenitor cells; Pulmonary; Pulmonary hypertension; Endothelium; Microfluidic device; Biomedical engineering; Biotechnology; Point-of-care testing; Endothelial cell; Stem cell; Lung; Biological marker; Cardiovascular disease; Chip; Respiratory system; Capture; Goal; Blood vessel; Human; Respiratory disease; Biological indicator; Patient; Artery; Test validation; Medical device; Circulatory system; Progenitor cell
• ISSN: 0946-2716; 1432-1440
• DOI: 10.1007/s00109-011-0779-6

The number of circulating endothelial progenitor cells (EPCs) inversely correlates with cardiovascular risk and clinical outcome, and thus has been proposed as a valuable biomarker for risk assessment, disease progression, and response to therapy. However, current strategies for isolation of these rare cells are limited to complex, laborious approaches. The goal of this study was the design and validation of a disposable microfluidic platform capable of selectively capturing and enumerating EPCs directly from human whole blood in healthy and diseased subjects, eliminating sample preprocessing. We then applied the “EPC capture chip” clinically and determined EPC numbers in blood from patients with pulmonary arterial hypertension (PAH). Blood was collected in tubes and injected into polymeric microfluidic chips containing microcolumns pre-coated with anti-CD34 antibody. Captured cells were immunofluorescently stained for the expression of stem and endothelial antigens, identified and counted. The EPC capture chip was validated with conventional flow cytometry counts ( r  = 0.83). The inter- and intra-day reliability of the microfluidic devices was confirmed at different time points in triplicates over 1–5 months. In a cohort of 43 patients with three forms of PAH (idiopathic/heritable, drug-induced, and connective tissue disease), EPC numbers are ≈50% lower in PAH subjects vs. matched controls and inversely related to two potential disease modifiers: body mass index and postmenopausal status. The EPC capture chip (5 × 30 × 0.05 mm 3 ) requires only 200 μL of human blood and has the strong potential to serve as a rapid bedside test for the screening and monitoring of patients with PAH and other proliferative cardiovascular, pulmonary, malignant, and neurodegenerative diseases.