title: Microfluidic assessment of functional culture-derived platelets in human thrombi under flow.
abstract: Despite their clinical significance, human platelets are not amenable to genetic manipulation, thus forcing a reliance on mouse models. Culture-derived platelets (CDPs) from human peripheral blood CD34(+) cells can be genetically altered and may eventually be used for transfusions. By use of microfluidics, the time-dependent incorporation of CD41(+)CD42(+) CDPs into clots was measured using only 54,000 CDPs doped into 27 μL of human whole blood perfused over collagen at a wall shear rate of 100 sec(-1). With the use of fluorescence-labeled human platelets (instead of CDPs) doped between 0.25% and 2% of total platelets, incorporation was highly quantitative and allowed monitoring of the anti-αIIbβ3 antagonism that occurred after collagen adhesion. CDPs were only 15% as efficient as human platelets in their incorporation into human thrombi under flow, although both cell types were equally antagonized by αIIbβ3 inhibition. Transient transfection allowed the monitoring of GFP(+) human CDP incorporation into clots. This assay quantifies genetically altered CDP function under flow.
authors.affiliation: Department of Chemical and Biomolecular Engineering, Institute for Medicine and Engineering, University of Pennsylvania, Philadelphia, Pennsylvania.
authors.forename: Viraj
authors.initials: V
authors.lastname: Kamat
authors.affiliation: Department of Chemical and Biomolecular Engineering, Institute for Medicine and Engineering, University of Pennsylvania, Philadelphia, Pennsylvania.
authors.forename: Ryan W
authors.initials: RW
authors.lastname: Muthard
authors.affiliation: Department of Chemical and Biomolecular Engineering, Institute for Medicine and Engineering, University of Pennsylvania, Philadelphia, Pennsylvania.
authors.forename: Ruizhi
authors.initials: R
authors.lastname: Li
authors.affiliation: Department of Chemical and Biomolecular Engineering, Institute for Medicine and Engineering, University of Pennsylvania, Philadelphia, Pennsylvania. Electronic address: sld@seas.upenn.edu.
authors.forename: Scott L
authors.initials: SL
authors.lastname: Diamond
mesh_terms: D000818:Animals; D015703:Antigens, CD / Q000378:metabolism; D001777:Blood Coagulation; D001792:Blood Platelets / Q000166:cytology / Q000378:metabolism*; D018929:Cell Culture Techniques / Q000379:methods; D002478:Cells, Cultured; D005260:Female; D006412:Hematopoietic Stem Cells / Q000166:cytology / Q000378:metabolism*; D006801:Humans; D008297:Male; D051379:Mice; D019039:Platelet Glycoprotein GPIIb-IIIa Complex / Q000378:metabolism
chemical_list: D015703:Antigens, CD; D019039:Platelet Glycoprotein GPIIb-IIIa Complex