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University of California San Francisco and University of California Berkeley

3 ARTICLES PUBLISHED IN JoVE

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Bioengineering

Patterning the Geometry of Human Embryonic Stem Cell Colonies on Compliant Substrates to Control Tissue-Level Mechanics
Jonathon M. Muncie 1,2, Roberto Falcón-Banchs 1, Johnathon N. Lakins 2, Lydia L. Sohn 1,3, Valerie M. Weaver 2,4,5,6
1Graduate Program in Bioengineering, University of California San Francisco and University of California Berkeley, 2Center for Bioengineering and Tissue Regeneration, Department of Surgery, University of California San Francisco, 3Department of Mechanical Engineering, University of California Berkeley, 4Eli and Edythe Broad Center of Regeneration Medicine and Stem Cell Research, University of California San Francisco, 5UCSF Comprehensive Cancer Center, Helen Diller Family Cancer Research Center, University of California San Francisco, 6Department of Anatomy, Department of Bioengineering and Therapeutic Sciences, and Department of Radiation Oncology, University of California San Francisco

Extracellular matrix ligands can be patterned onto polyacrylamide hydrogels to enable the culture of human embryonic stem cells in confined colonies on compliant substrates. This method can be combined with traction force microscopy and biochemical assays to examine the interplay between tissue geometry, cell-generated forces, and fate specification.

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Bioengineering

Simple, Affordable, and Modular Patterning of Cells using DNA
Katelyn A. Cabral 1, David M. Patterson 2, Olivia J. Scheideler 1, Russell Cole 3, Adam R. Abate 4,5,6, David V. Schaffer 7,8, Lydia L. Sohn 9, Zev J. Gartner 2,6,10
1Graduate Program in Bioengineering, University of California San Francisco and University of California Berkeley, 2Department of Pharmaceutical Chemistry, University of California San Francisco, 3Scribe Biosciences, 4Department of Bioengineering and Therapeutic Sciences, University of California San Francisco, 5California Institute for Quantitative Biosciences, University of California San Francisco, 6Chan Zuckerberg Biohub, University of California San Francisco, 7Department of Chemical & Biomolecular Engineering, University of California Berkeley, 8Helen Wills Neuroscience Institute, University of California Berkeley, 9Department of Mechanical Engineering, University of California Berkeley, 10Center for Cellular Construction, University of California San Francisco

Here we present a protocol to micropattern cells at single-cell resolution using DNA-programmed adhesion. This protocol uses a benchtop photolithography platform to create patterns of DNA oligonucleotides on a glass slide and then labels cell membranes with commercially available complementary oligonucleotides. Hybridization of the oligos results in programmed cell adhesion.

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Bioengineering

Mechano-Node-Pore Sensing: A Rapid, Label-Free Platform for Multi-Parameter Single-Cell Viscoelastic Measurements
Andre Lai *1, Rachel Rex *2, Kristen L. Cotner 1, Alan Dong 3, Michael Lustig 1,3, Lydia L. Sohn 1,2
1Graduate Program in Bioengineering, University of California, Berkeley and University of California, San Francisco, 2Department of Mechanical Engineering, University of California, Berkeley, 3Department of Electrical Engineering and Computer Sciences, University of California, Berkeley

Presented here is a method to mechanically phenotype single cells using an electronics-based microfluidic platform called mechano-node-pore sensing (mechano-NPS). This platform maintains moderate throughput of 1-10 cells/s while measuring both the elastic and viscous biophysical properties of cells.

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