Novel Low Shear 3D Bioreactor for the Scaled Production of High Purity Human Mesenchymal Stem Cells

Author

Andrew BurnsFollow

Date Degree Awarded

Spring 5-18-2019

Degree Type

Restricted to Claremont Colleges Dissertation

Degree Name

PHD in Applied Life Sciences

First Thesis/Dissertation Advisor

M. Ian Phillips

Second Thesis/Dissertation Advisor

Parviz Shamlou

Third Thesis/Dissertation Advisor

Cameron Bardliving

Terms of Use & License Information

Terms of Use for work posted in Scholarship@Claremont.

Abstract

Human mesenchymal stem cells are an ideal candidate for stem cell therapies. They have been researched since the 1960’s and can differentiate into many desired functional cell types without undergoing teratogenesis. However, higher yields are needed for a marketable, successful stem cell therapy. To accomplish this, cells will have to be cultured to expand them to therapeutically relevant dosages for multiple patients. Bioreactor production is an ideal method to solve this problem.

The aim of this thesis is to test and validate a novel bioreactor for the cultivation of human mesenchymal stem cells. In this work, we investigate a novel suspended matrix for the culture on human mesenchymal stem cells (hMSCs). Initially we investigated various fiber meshes, both random and structured, for stem cell growth and morphology. We also investigated hMSC proliferation on rigid polymers commonly used in 3D printing. We then took the conditions that worked best in 2D culture and tested them in a small-scale model of the Express bioreactor from Sepragen.

We have assessed cell growth on 3D printed Polylactic Acid (PLA) matrices and developed a scale down model bioreactor for development and characterization. Computational Fluid Dynamic (CFD) modeling was used in parallel with the described in-vitro experimentation to characterize shear profiles. From the CFD we were also able to predict a flow rate which resulted in almost zero shear. What we found was that hMSCs readily form confluent monolayers on the PLA lattice, and retain their surface marker expression and stemness. When combined with a short hypoxic treatment, the cells performed better than control flasks, resulting in a four-fold increase from seed with no impact on biomarker profile and differentiation ability.

Rights Information

© 2019 Andrew B Burns

Recommended Citation

Burns, Andrew. (2019). Novel Low Shear 3D Bioreactor for the Scaled Production of High Purity Human Mesenchymal Stem Cells. KGI Theses and Dissertations, 21. https://scholarship.claremont.edu/kgi__theses/21.