The ability to harness the processes by which complex tissues arise during embryonic development would improve the ability to engineer complex tissuelike constructs in vitro—a longstanding goal of tissue engineering and regenerative medicine. In embryos, uniform populations of stem cells are exposed to spatial gradients of diffusible extracellular signaling proteins, known as morphogens. Varying levels of these signaling proteins induce stem cells to differentiate into distinct cell types at different positions along the gradient, thus creating spatially patterned tissues. Here, the authors describe two straightforward and easy-to-adopt microfluidic strategies to expose human pluripotent stem cells in vitro to spatial gradients of desired differentiation-inducing extracellular signals. Both approaches afford a high degree of control over the distribution of extracellular signals, while preserving the viability of the cultured stem cells. The first microfluidic platform is commercially available and entails static culture, whereas the second microfluidic platform requires fabrication and dynamic fluid exchange. In each platform, the authors first computationally modeled the spatial distribution of differentiation-inducing extracellular signals. Then, the authors used each platform to expose human pluripotent stem cells to a gradient of these signals (in this case, inducing a cell type known as the primitive streak), resulting in a regionalized culture with differentiated primitive streak cells predominately localized on one side and undifferentiated stem cells at the other side of the device. By combining this approach with a fluorescent reporter for differentiated cells and live-cell fluorescence imaging, the authors characterized the spatial and temporal dynamics of primitive streak differentiation within the induced signaling gradients. Microfluidic approaches to create precisely controlled morphogen gradients will add to the stem cell and developmental biology toolkit, and may eventually pave the way to create increasingly spatially patterned tissuelike constructs in vitro.
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Research Article|
March 24 2020
Spatially controlled stem cell differentiation via morphogen gradients: A comparison of static and dynamic microfluidic platforms
Kiara W. Cui
;
Kiara W. Cui
1
Department of Chemical Engineering, Stanford University
, Stanford, California 94305
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Leeya Engel
;
Leeya Engel
1
Department of Chemical Engineering, Stanford University
, Stanford, California 94305
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Carolyn E. Dundes;
Carolyn E. Dundes
2
Department of Developmental Biology, Stanford Institute for Stem Cell Biology and Regenerative Medicine, Stanford University, Stanford
, California 94305
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Tina C. Nguyen;
Tina C. Nguyen
1
Department of Chemical Engineering, Stanford University
, Stanford, California 94305
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Kyle M. Loh
;
Kyle M. Loh
2
Department of Developmental Biology, Stanford Institute for Stem Cell Biology and Regenerative Medicine, Stanford University, Stanford
, California 94305
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Alexander R. Dunn
Alexander R. Dunn
d)
1
Department of Chemical Engineering, Stanford University
, Stanford, California 94305
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a)
K. W. C. and L. E. contributed equally to this work.
b)
Electronic mail: kiaracui@stanford.edu
c)
Electronic mail: leeya@stanford.edu
d)
Authors to whom correspondence should be addressed: kyleloh@stanford.edu and alex.dunn@stanford.edu
e)
Electronic mail: kyleloh@stanford.edu
Note: This paper is part of the Special Topic Collection on 30 years of the Nellie Yeoh Whetten Award — Celebrating the Women of the AVS.
J. Vac. Sci. Technol. A 38, 033205 (2020)
Article history
Received:
December 09 2019
Accepted:
February 21 2020
Citation
Kiara W. Cui, Leeya Engel, Carolyn E. Dundes, Tina C. Nguyen, Kyle M. Loh, Alexander R. Dunn; Spatially controlled stem cell differentiation via morphogen gradients: A comparison of static and dynamic microfluidic platforms. J. Vac. Sci. Technol. A 1 May 2020; 38 (3): 033205. https://doi.org/10.1116/1.5142012
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