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.

Cell separation using tilted-angle standing surface acoustic waves

“…When separating cells with distinct physical properties, using methods that exploit differences in cells' physical parameters could be advantageous due to their label-free nature and ease of use (11,12). Many techniques are available to separate cells based on physical properties; they include filtration, centrifugation, acoustics, optics, and dielectrophoresis (13)(14)(15)(16)(17)(18)(19)(20)(21)(22)(23)(24)(25)(26)(27). Among these techniques, acoustic-based approaches are advantageous because of their biocompatibility and label-free nature (25,(27)(28)(29)(30)(31)(32).…”

Micro Total Analysis Systems: Fundamental Advances and Applications in the Laboratory, Clinic, and Field

“…Another group purified hematopoietic stem cells from harvested bone marrow for regenerative medicine using a PDMS filter that achieved greater than 90% enrichment and post-separation viability. 218 …”

Selective Cell Recruitment and Spatially Controlled Cell Attachment on Instructive Chitosan Surfaces Functionalized with Antibodies

“…Compared with currently available devices found in literature the system herein proposed besides effective it is relatively low cost using a biomaterial with easy chemistry that can be later implantable [10][11][12][13][14][15][16]. The method proposed in this work allows the isolation of a target cell type from a mixed cell population with relatively high purity, decreasing cell handling where recruited cells can be used straightaway.…”

“…Cell sorting according to their size has been proposed as an effective and rapid method to separate cells [10,11]. Dielectrophoresis separates cells according to their electric properties and/or size [12].…”