title: Capture and enrichment of CD34-positive haematopoietic stem and progenitor cells from blood circulation using P-selectin in an implantable device.
abstract: Clinical infusion of haematopoietic stem and progenitor cells (HSPCs) is vital for restoration of haematopoietic function in many cancer patients. Previously, we have demonstrated an ability to mimic physiological cell trafficking in order to capture CD34-positive (CD34+) HSPCs using monolayers of the cell adhesion protein P-selectin in flow chambers. The current study aimed to determine if HSPCs could be captured directly from circulating blood in vivo. Vascular shunt prototypes, coated internally with P-selectin, were inserted into the femoral artery of rats. Blood flow through the cell capture device resulted in a wall shear stress of 4-6 dynes/cm(2). After 1-h blood perfusion, immunofluorescence microscopy and flow cytometric analysis revealed successful capture of mononuclear cells positive for the HSPC surface marker CD34. Purity of captured CD34+ cells showed sevenfold enrichment over levels found in whole blood, with an average purity of 28%. Robust cell capture and HSPC enrichment were also demonstrated in devices that were implanted in a closed-loop arterio-venous shunt conformation for 2 h. Adherent cells were viable in culture and able to differentiate into burst-forming units. This study demonstrated an ability to mimic the physiological arrest of HSPCs from blood in an implantable device and may represent a practical alternative for adult stem cell capture and enrichment.
authors.affiliation: Department of Biomedical Engineering, University of Rochester, Rochester, NY 14627, USA.
authors.forename: Joel C
authors.initials: JC
authors.lastname: Wojciechowski
authors.forename: Srinivas D
authors.initials: SD
authors.lastname: Narasipura
authors.forename: Nichola
authors.initials: N
authors.lastname: Charles
authors.forename: Deanne
authors.initials: D
authors.lastname: Mickelsen
authors.forename: Kuldeeepsinh
authors.initials: K
authors.lastname: Rana
authors.forename: Martha L
authors.initials: ML
authors.lastname: Blair
authors.forename: Michael R
authors.initials: MR
authors.lastname: King
mesh_terms: D000818:Animals; D018952:Antigens, CD34 / Q000097:blood*; D002448:Cell Adhesion; D002455:Cell Division; D002469:Cell Separation / Q000379:methods*; D002470:Cell Survival; D003114:Colony-Forming Units Assay; D005263:Femoral Artery; D005434:Flow Cytometry; D006412:Hematopoietic Stem Cells / Q000166:cytology* / Q000276:immunology / Q000378:metabolism; D020007:Implants, Experimental; D008297:Male; D019007:P-Selectin / Q000378:metabolism*; D051381:Rats; D017207:Rats, Sprague-Dawley
chemical_list: D018952:Antigens, CD34; D019007:P-Selectin