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Full text release has been delayed at the author's request until June 01, 2025

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3D EXPANSION AND DIFFERENTIATION OF INDUCED PLURIPOTENT STEM CELLS INTO INSULIN-PRODUCING β-CELLS

Yehya, Haneen Samer
http://orcid.org/0009-0006-6117-7442
http://rave.ohiolink.edu/etdc/view?acc_num=csu1715346647428199

Abstract Details

2024, Doctor of Philosophy in Engineering, Cleveland State University, Washkewicz College of Engineering.
Cell therapies derived from induced pluripotent stem cells iPSCs hold promise in a wide array of clinical areas due to their capacity to differentiate into various cell types. Among these, pancreatic iPSC derived β-cells for treating Type 1 diabetes. However, there are many challenges surrounding scaling iPSC production and their derivatives such as β- cells for clinical use. These challenges have been addressed by developing an optimized bioengineered process for expanding iPSCs and differentiating them into β-cells. Through comprehensive experimentation and mathematical modeling, this research optimizes iPSC production, maintenance, aggregation, and pancreatic differentiation process and protocol transfer into a 3D environment, by defining critical process parameters (CPPs) and critical quality attributes (CQAs) essential for clinical translation. The goal of this research work is to contribute to the clinical translation of iPSC- based therapies and specifically for Type 1 diabetes by leveraging multifactorial perturbation methodology and bioprocess understanding to generate a scale necessary for clinical treatments. The study has two main focuses. One is to optimize the cell medium needed for iPSC maintenance and aggregate stability utilizing a design of experiments (DoE) approach in PBS vertical wheel bioreactors. The second focus is on the specific differentiation of iPSCs into β-cells, employing an optimized HD-DoE protocol and designing it for scalability in a suspension environment implemented in vertical wheel bioreactors with process optimized parameters from seeding to banking. The iPSC expansion, maintenance and optimization in 100 ml vertical wheel bioreactors tested in a factorial design identifies combinational media additives that attain pluripotency, continuous growth, and aggregate stability. The three-stage suspension pancreatic differentiation protocol developed for β-cells, is compared to an adherent differentiation then it transitions into bioreactor systemic optimization that achieves enhanced marker expression and differentiation of iPSC-derived islet-like clusters. Constant medium bioreactor runs with no media replenishment are studied for better process understanding. Their differentiation and metabolic profiles are compared to control bioreactors receiving frequent media changes between stages. Overall, this research offers insights about media optimization strategy for complex biological processes and systemic approach for addressing bioreactor scale-up challenges, highlighting the potential of iPSC-derivatives as being sources of widespread clinical therapy.
Michael Bukys (Advisor)
193 p.
Biomedical Engineering; Biomedical Research; Chemical Engineering; Engineering
Diabetes; human induced pluripotent stem cell; Insulin Producing Cells; Bioreactor; DoE; β-cells; Pancreatic cells; bioprocess development; optimization; islets; iPSCs; manufacturing; aggregates; growth; suspension; 3D culture; Type I diabetes

Recommended Citations

Citations

  • Yehya, H. S. (2024). 3D EXPANSION AND DIFFERENTIATION OF INDUCED PLURIPOTENT STEM CELLS INTO INSULIN-PRODUCING β-CELLS [Doctoral dissertation, Cleveland State University]. OhioLINK Electronic Theses and Dissertations Center. http://rave.ohiolink.edu/etdc/view?acc_num=csu1715346647428199

    APA Style (7th edition)

  • Yehya, Haneen. 3D EXPANSION AND DIFFERENTIATION OF INDUCED PLURIPOTENT STEM CELLS INTO INSULIN-PRODUCING β-CELLS. 2024. Cleveland State University, Doctoral dissertation. OhioLINK Electronic Theses and Dissertations Center, http://rave.ohiolink.edu/etdc/view?acc_num=csu1715346647428199.

    MLA Style (8th edition)

  • Yehya, Haneen. "3D EXPANSION AND DIFFERENTIATION OF INDUCED PLURIPOTENT STEM CELLS INTO INSULIN-PRODUCING β-CELLS." Doctoral dissertation, Cleveland State University, 2024. http://rave.ohiolink.edu/etdc/view?acc_num=csu1715346647428199

    Chicago Manual of Style (17th edition)

csu1715346647428199
© 2024, all rights reserved.
This open access ETD is published by Cleveland State University and OhioLINK.
Release 3.2.12