Date of Award

5-2024

Document Type

Thesis

Degree Name

Master of Science in Biology

Department

Biology

First Reader/Committee Chair

Bournias-Vardiabasis, Nicole

Abstract

Neural stem cell therapies represent a promising tool for the development of regenerative medicine and are being tested in clinical trials for several neurological disorders. However, the clinical applicability of stem cell therapies is dependent on the implementation of current good manufacturing practices (cGMPs) to ensure the quality, safety, and consistency that stem cell products need to meet FDA regulatory requirements. As such, there is a need for a shift to xeno-free methodologies so experimental conditions are cGMP compliant. The purpose of this study is to test a GMP compatible production method to generate multipotent neural stem cells (NSCs) from embryonic and pluripotent stem cells. Comparability of NSCs is dependent on quality controls such as safety, stability, purity, and multipotent neural stem cell differentiation. Accordingly, we will test Shef6 hESC and ADRC76 hiPSC lines for their suitability to produce CD133-positive neural stem cells. Due to potential epigenetic differences found in hiPSCs, such as aberrations in DNA, we predict that hESCs will produce higher quality NSCs compared to hiPSCs. To test this, both cell lines will be cultured under identical reagents and methods for expansion and neuralization. For NSC characterization, trophic factor and cytokine secretion and neural differentiation will be compared between the processing runs. General Biological Product Standards will be applied throughout the downstream processing such as safety, stability, potency and purity validations. This manufacturing protocol will be compared with a prior derivation of Shef6 hNSCs. The adoption of cGMP-compliant methods early in the research environment will improve the replication of results across cell lines and may increase translatability of preclinical studies as the protocols are transferred to GMP facilities.

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