Cell seeding and proliferation simulations to plan experiments of multicellular osteogenic systems

Osteogenesis in vivo involves changes to cell density, morphology, cell-cell contacts, gene expression, and proliferation; however, the coordination of these changes is unknown. Specifically, cells undergo a morphological switch; they are proliferative, pluripotent, elongated cells early in differentiation and become densely packed, cuboidal, mature osteoblasts. Osteogenic differentiation of stem cells can be promoted in vitro via geometry using micropatterned substrates, or via seeding densities that allow for cell spreading. Due to the complexity of biological systems, typically, researchers use patterned substrates and inhibit proliferation, or observe proliferation on unpatterned substrates to isolate and understand the influence of each factor. However, these limitations impact results. In this work, to examine proliferation on patterned islands, we create, validate, and demonstrate use of cell seeding and proliferation simulations on micropatterned islands. These simulations mirror experiments and provide more data than possible from experiments alone. We can use these simulations to plan experiments to examine cell behaviors. We show that these cell behaviors can be predicted by confluence, which incorporates time in culture, the current standard means for comparison, seeding density, and island size. Using confluence as a standard for comparison reduces variability in experimental methods between researchers. We also show that on islands with area to perimeter ratio greater than the cell size, cells behave more like a bulk monolayer of cells; this work is the first to suggest the importance of island area to perimeter ratio. As demonstrated by this work, these simulations can be used as hypothesis generating tool or a tool to plan experiments to achieve a desired range of confluence based on island geometry, seeding density, and time in culture.

Name thesis:degree_name

Ph.D.

Level thesis:degree_level

Dissertation

Discipline thesis:degree_discipline

Mechanical Engineering

Grantor

University of Illinois at Urbana-Champaign

Year dc:date

2021

Author dc:creator

• Berent, Zachary T

Contributors dc:contributor

• Wagoner Johnson, Amy
• Harley, Brendan
• Underhill, Gregory
• Kersh, Mariana

• osteogenesis, mesenchymal stem cell, micropatterning

Statement dc:rights

• Copyright 2020 Zachary Berent

Language dc:language

en

Handle dc:identifier

http://hdl.handle.net/2142/109593

OAI identifier oai:identifier

oai:www.ideals.illinois.edu:2142/109593