Stem cells have traditionally served as a valuable resource for models of development and morphogenesis, but they also factor significantly in the development of regenerative therapies aiming to treat various degenerative diseases and traumatic injuries. Pluripotent embryonic stem cells (ESCs) are capable of differentiating into an array of cell types, including functional neurons, cardiomyocytes and pancreatic beta cells, thus representing a robust cell source for biological studies and regenerative cell therapies. Despite the clear potential of pluripotent stem cells, a critical limitation is the inability to efficiently differentiate ESCs to specific cell fates in a reproducible, reliable and homogeneous manner. During embryonic development, the spatiotemporal fidelity of differentiation is precisely regulated by the local amount of morphogenic factors presented at the appropriate time with a finite duration. However, most in vitro methods typically used to examine the differentiation of stem cells in response to morphogen treatment lack the ability to uniformly present molecules in defined amounts with precise temporal resolution. Forced convection intercellular perfusion (FCIP) created by microfluidic culture systems can overcome the limitations of diffusive transport by presenting molecules to 3D cultures of cells in a uniform temporal and dose-dependent controlled manner. The inherent flexibility and scalability of microfluidic perfusion culture systems represents an innovative and translatable approach to enhance directed stem cell differentiation. Based on this rationale and significant preliminary data, the objectives of this proposal are to 1) define the transport limitations of 3D aggregates of differentiating ESCs, 2) determine the effects of perfusion culture on the yield and homogeneity of ESC differentiation, and 3) examine the dose and temporal effects of morphogen presentation on ESC differentiation in a high-throughput manner. The completion of these studies will yield novel information about the ability to more controllably direct the differentiation of ESCs by engineering the dynamic molecular composition of the extracellular microenvironment using microfluidic perfusion culture. The proposed approach represents a fundamentally new route to more efficiently direct the differentiation of stem cells in vitro through the simultaneous control of dose and temporal presentation of molecular factors locally, which may be a broadly applicable principle in the development of stem cell technologies.
The development of stem cell regenerative and diagnostic technologies is currently limited by an inability to efficiently control the differentiation of the stem cells. This proposal seeks to improve the efficiency and homogeneity of pluripotent stem cell differentiation by controlling the dose and timing of morphogenic factor presentation within 3D stem cell microenvironments via microperfusion culture systems.
|White, Douglas E; Sylvester, Jonathan B; Levario, Thomas J et al. (2015) Quantitative multivariate analysis of dynamic multicellular morphogenic trajectories. Integr Biol (Camb) 7:825-33|
|Wilson, Jenna L; Suri, Shalu; Singh, Ankur et al. (2014) Single-cell analysis of embryoid body heterogeneity using microfluidic trapping array. Biomed Microdevices 16:79-90|
|Kinney, Melissa A; Saeed, Rabbia; McDevitt, Todd C (2014) Mesenchymal morphogenesis of embryonic stem cells dynamically modulates the biophysical microtissue niche. Sci Rep 4:4290|
|Kinney, Melissa A; Hookway, Tracy A; Wang, Yun et al. (2014) Engineering three-dimensional stem cell morphogenesis for the development of tissue models and scalable regenerative therapeutics. Ann Biomed Eng 42:352-67|
|Wilson, Jenna L; Najia, Mohamad Ali; Saeed, Rabbia et al. (2014) Alginate encapsulation parameters influence the differentiation of microencapsulated embryonic stem cell aggregates. Biotechnol Bioeng 111:618-31|
|Kinney, Melissa A; Sargent, Carolyn Y; McDevitt, Todd C (2013) Temporal modulation of *-catenin signaling by multicellular aggregation kinetics impacts embryonic stem cell cardiomyogenesis. Stem Cells Dev 22:2665-77|
|Kinney, Melissa A; McDevitt, Todd C (2013) Emerging strategies for spatiotemporal control of stem cell fate and morphogenesis. Trends Biotechnol 31:78-84|
|White, Douglas E; Kinney, Melissa A; McDevitt, Todd C et al. (2013) Spatial pattern dynamics of 3D stem cell loss of pluripotency via rules-based computational modeling. PLoS Comput Biol 9:e1002952|
|Suri, Shalu; Singh, Ankur; Nguyen, Anh H et al. (2013) Microfluidic-based patterning of embryonic stem cells for in vitro development studies. Lab Chip 13:4617-24|
|Wilson, Jenna L; McDevitt, Todd C (2013) Stem cell microencapsulation for phenotypic control, bioprocessing, and transplantation. Biotechnol Bioeng 110:667-82|
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