Suspension culture is the gold standard for ex vivo hematopoietic stem cell (HSC) expansion; however, high media requirement results in expensive cell therapy products. Hydrogel-based HSC culture represents a more realistic in vitro representation of human in vivo HSC microenvironments and may support a more efficient use of media during ex vivo HSC expansion. However, media conditions used for hydrogel-supported cell culture often simply mimic that which are currently used in suspension culture approaches, potentially resulting in an oversupply of cell growth proteins. In this work, we present a microfluidic culture system capable of supporting hydrogel-based cell culture and generating spatial distributions of independent growth factor combinations. Using this device, we image, in real-time, the behaviour of human umbilical cord blood-derived CD34+ cells, within fibrin-based hydrogels, in response to spatial gradients of the human recombinant growth factors stem cell factor, thrombopoietin, angiopoietin-2 and insulin-like growth factor-II. Using live microscopy and image-based single-cell segmentation and quantification, local changes in cell density over time in relation to spatial concentration of factors were identified. This novel microfluidic device has the potential to screen combinations of growth factors required for hydrogel-supported CD34+ cell expansion and inform culture media formulations of future hydrogel-based cell manufacturing processes.
Murphy, A. R., Franco, R. A., Johnson, P., Cooper-White, J. J., Allenby, M. C.
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