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Issue 12, 2013
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Robust pluripotent stem cell expansion and cardiomyocyte differentiation via geometric patterning

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Abstract

Geometric factors including the size, shape, density, and spacing of pluripotent stem cell colonies play a significant role in the maintenance of pluripotency and in cell fate determination. These factors are impossible to control using standard tissue culture methods. As such, there can be substantial batch-to-batch variability in cell line maintenance and differentiation yield. Here, we demonstrate a simple, robust technique for pluripotent stem cell expansion and cardiomyocyte differentiation by patterning cell colonies with a silicone stencil. We have observed that patterning human induced pluripotent stem cell (hiPSC) colonies improves the uniformity and repeatability of their size, density, and shape. Uniformity of colony geometry leads to improved homogeneity in the expression of pluripotency markers SSEA4 and Nanog as compared with conventional clump passaging. Patterned cell colonies are capable of undergoing directed differentiation into spontaneously beating cardiomyocyte clusters with improved yield and repeatability over unpatterned cultures seeded either as cell clumps or uniform single cell suspensions. Circular patterns result in a highly repeatable 3D ring-shaped band of cardiomyocytes which electrically couple and lead to propagating contraction waves around the ring. Because of these advantages, geometrically patterning stem cells using stencils may offer greater repeatability from batch-to-batch and person-to-person, an increase in differentiation yield, a faster experimental workflow, and a simpler protocol to communicate and follow. Furthermore, the ability to control where cardiomyocytes arise across a culture well during differentiation could greatly aid the design of electrophysiological assays for drug-screening.

Graphical abstract: Robust pluripotent stem cell expansion and cardiomyocyte differentiation via geometric patterning

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Publication details

The article was received on 12 Aug 2012, accepted on 08 Oct 2013, published on 10 Oct 2013 and first published online on 10 Oct 2013


Article type: Technical Innovation
DOI: 10.1039/C2IB20191G
Citation: Integr. Biol., 2013,5, 1495-1506
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    Robust pluripotent stem cell expansion and cardiomyocyte differentiation via geometric patterning

    F. B. Myers, J. S. Silver, Y. Zhuge, R. E. Beygui, C. K. Zarins, L. P. Lee and O. J. Abilez, Integr. Biol., 2013, 5, 1495
    DOI: 10.1039/C2IB20191G

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