title: Microfluidic purification and analysis of hematopoietic stem cells from bone marrow.
abstract: Hematopoietic stem cells are larger in size than other cells present in bone marrow, with the exception of monocytes. This distinguishing characteristic can be used to separate them from a whole-marrow sample. A microfluidic device was fabricated using an integrated membrane that is porous at defined areas. This allows for simultaneous valving and filtering functionality, which is crucial for preventing irreversible clogging. This device, as well as a separation procedure, was optimized in this work to enrich hematopoietic progenitor cells from diluted bone marrow of leukemia patients without any additional sample preparation. An enrichment of up to 98% was achieved with this method and the process was scaled up to 17.2 μL min(-1) of processed sample. Additionally, stem cells were stained with specific antibodies for further analysis. Using a custom-made computer program, the filter was scanned to characterize and quantify cells based on fluorescence. The results were evaluated by comparing them against the results obtained from flow cytometry, confocal microscopy, and Coulter counting.
authors.affiliation: Department of Chemistry, Stanford University, Stanford, California 94305-5080, USA.
authors.forename: Romana
authors.initials: R
authors.lastname: Schirhagl
authors.forename: Ingo
authors.initials: I
authors.lastname: Fuereder
authors.forename: Eric W
authors.initials: EW
authors.lastname: Hall
authors.forename: Bruno C
authors.initials: BC
authors.lastname: Medeiros
authors.forename: Richard N
authors.initials: RN
authors.lastname: Zare
mesh_terms: D000208:Acute Disease; D001854:Bone Marrow Cells / Q000166:cytology* / Q000473:pathology; D002469:Cell Separation / Q000295:instrumentation* / Q000379:methods; D048429:Cell Size; D002470:Cell Survival; D005434:Flow Cytometry; D005452:Fluoresceins / Q000737:chemistry; D006412:Hematopoietic Stem Cells / Q000166:cytology*; D006801:Humans; D007938:Leukemia / Q000473:pathology; D046210:Microfluidic Analytical Techniques / Q000295:instrumentation*; D018613:Microscopy, Confocal; D016062:Porosity
chemical_list: D005452:Fluoresceins