Titmarsh, D.1, Glass, N.1, Hudson, J.2, Hidalgo, A.1, Porrello, E.2, Wolvetang, E.1, Little, M.3 and Cooper-White, J.1 *
- Australian Institute for Bioengineering and Nanotechnology, The University of Queensland, St. Lucia, Brisbane, QLD 4072, Australia.
- Cardiac Regeneration Lab, School of Biomedical Sciences, The University of Queensland, St. Lucia, Brisbane, QLD 4072, Australia.
- Kidney Disease, Repair and Regeneration Lab, Murdoch Children’s Institute, Royal Children’s Hospital, Melbourne, VIC 3052, Australia.
* corresponding author - email@example.com
Deploying human stem cells (pluripotent (hPSCs) or mesenchymal (hMSCs)) in regenerative medicine depends on effective control of both their undifferentiated expansion and differentiation into desired lineages. To efficiently produce sufficient, defined cell populations, we must be able to direct stem cell fate choices, yet this is substantially hindered by undefined culture components, signal crosstalk between multiple exogenous and endogenous factors, and spatiotemporal variations in microenvironmental composition inherent to conventional culture formats. We have developed credit card sized, scalable, valveless, continuous-flow microbioreactor arrays (MBAs) that both provide a combinatorial set of exogenous factor compositions, and allow controlled accumulation of paracrine factors. These arrays have been used to survey up to 8100 individual, perfused cellular microenvironments in parallel. Through screens of pluripotency maintenance and differentiation of hPSCs into primitive streak, cardiac and kidney cells, as examples, we demonstrate the unique ability of this platform to separate, visualise, identify and modulate paracrine effects that are not otherwise readily accessible with standard culture formats. These MBAs have also been utilized to optimize media formulations for the expansion and osteogenic differentiation of hMSCs. Culture conditions optimized with the arrays have been shown to readily translate to conventional static culture protocols, leading to improved cardiac, kidney and bone differentiation. With our latest generation 8100-chamber MBA we have assessed the impacts and interplay of developmental factors on proliferation of hPSC-derived cardiomyocytes, exemplifying the utility of the device for patient-specific early drug stratification. These multiplexed MBA platforms decipher factor interplay and signalling hierarchies that control stem cell fate, and are applicable as a universal microenvironmental screening platform for bioprocess optimisation, media formulation design, quality control for cellular therapies and cell-based drug screening.