Engineering pluripotent stem cell heterogeneity and its effects on differentiation

Wang, Jason Xiao

Permalink

https://hdl.handle.net/2142/124561 Copy

Description

Title

Engineering pluripotent stem cell heterogeneity and its effects on differentiation

Author(s)

Wang, Jason Xiao

Issue Date

2024-04-26

Director of Research (if dissertation) or Advisor (if thesis)

• Kong, Hyunjoon
• Dar, Roy

Doctoral Committee Chair(s)

Kong, Hyunjoon

Committee Member(s)

• Underhill, Gregory
• Golding, Ido

Department of Study

Bioengineering

Discipline

Bioengineering

Degree Granting Institution

University of Illinois at Urbana-Champaign

Degree Name

Ph.D.

Degree Level

Dissertation

Keyword(s)

• Stem Cell
• Heterogeneity
• Stochastic Gene Expression
• Differentiation
• Cardiac Tissue
• Motor Neuron
• Small Molecule Inhibitors

Language

eng

Abstract

Pluripotent stem cells (PSCs) have the potential to revolutionize both fundamental and applied bioscience studies, yet their applications have been limited due to their heterogeneity. Stem cell heterogeneity can lead to several challenges, such as differentiation into undesired cell types, compromised functionality, and the growth of cancerous tumors. Therefore, despite nearly a century of stem cell research, the FDA has only approved hematopoietic stem cells for therapeutic applications. It is essential to understand and regulate the mechanism of heterogeneity for enhanced stem cell engineering. To this end, we developed and optimized a reliable and cost-effective method using flow cytometry, allowing us to analyze multiple samples with various treatments at the single-cell level for population heterogeneity. With the established quantification methodology, we implemented a heuristic approach to identify potential mechanisms that may influence heterogeneity. The screened mechanisms spanned a broad range of disciplines, from modulating the growth substrate's stiffness to inhibiting DNA gene expression regulatory mechanisms to controlling cell properties such as cell cycle. We are the first to demonstrate that modulation of the cell cycle could induce a consistent and significant effect on the pluripotent gene expression. PSCs with modulated cell cycles were then differentiated into cardiac and motor neuron lineages. We present a novel finding: partial cell cycle arrest, compared to untreated and near-complete arrest conditions, significantly enhanced the functionality of differentiated tissues. In addition to demonstrating a novel methodology for enhancing stem cell differentiation, these findings suggest that much remains to be understood about how heterogeneity influences differentiation. The work presented lays a foundational methodology for further investigating and engineering pluripotent heterogeneity and investigating its effects on differentiation.

Graduation Semester

2024-05

Type of Resource

Text

Handle URL

https://hdl.handle.net/2142/124561

Copyright and License Information

Copyright 2024 Jason Wang

Owning Collections