Engineering Biophysical Cues for Controlled 3D Differentiation of Endoderm Derivatives

Thomas Richardson; Shibin Mathew; Connor Wiegand; Kevin Pietz; Joseph Candiello; K Ravikumar; Ipsita Banerjee

doi:10.1007/978-1-0716-1174-6_6

Engineering Biophysical Cues for Controlled 3D Differentiation of Endoderm Derivatives

Methods Mol Biol. 2021:2258:73-92. doi: 10.1007/978-1-0716-1174-6_6.

Authors

Thomas Richardson^{1

2}, Shibin Mathew^{1

3}, Connor Wiegand¹, Kevin Pietz⁴, Joseph Candiello^{4

5}, K Ravikumar¹, Ipsita Banerjee^{6

7

8}

Affiliations

¹ Department of Chemical and Petroleum Engineering, University of Pittsburgh, Pittsburgh, PA, USA.
² Lonza, Houston, Texas, PA, USA.
³ Pfiezer, Boston, MA, USA.
⁴ Department of Bioengineering, University of Pittsburgh, Pittsburgh, PA, USA.
⁵ RoosterBio, Frederick, MD, USA.
⁶ Department of Chemical and Petroleum Engineering, University of Pittsburgh, Pittsburgh, PA, USA. ipb1@pitt.edu.
⁷ Department of Bioengineering, University of Pittsburgh, Pittsburgh, PA, USA. ipb1@pitt.edu.
⁸ McGowan Institute for Regenerative Medicine, University of Pittsburgh School of Medicine, Pittsburgh, PA, USA. ipb1@pitt.edu.

PMID: 33340355
DOI: 10.1007/978-1-0716-1174-6_6

Abstract

Biophysical cues synergize with biochemical cues to drive differentiation of pluripotent stem cells through specific phenotypic trajectory. Tools to manipulate the cell biophysical environment and identify the influence of specific environment perturbation in the presence of combinatorial inputs will be critical to control the development trajectory. Here we describe the procedure to perturb biophysical environment of pluripotent stem cells while maintaining them in 3D culture configuration. We also discuss a high-throughput platform for combinatorial perturbation of the cell microenvironment, and detail a statistical procedure to extract dominant environmental influences.

Keywords: 3D Cell culture; Alginate encapsulation; Alginate stiffness; Biophysical cues; hPSC differentiation.

MeSH terms

Alginates / chemistry
Cell Culture Techniques
Cell Differentiation*
Cell Lineage*
Cells, Cultured
Endoderm / cytology
Endoderm / physiology*
Fluorescent Antibody Technique*
Gene Expression Regulation, Developmental
Humans
Mechanotransduction, Cellular*
Microscopy, Fluorescence*
Models, Statistical
Phenotype
Pluripotent Stem Cells / physiology*
Stem Cell Niche*
Time Factors
Tissue Engineering*

Substances

Alginates