Development of Cell Culture Models for Multiple Myeloma Drug Discovery: Comparison of Traditional 2D, Transwell 2D and 3D-Bioprinted Models
Date of Award
5-2024
Degree Type
Thesis
Degree Name
Master of Science in Biomedical Sciences
First Advisor
Xinyu (Eric) Wang
Second Advisor
Harold Komiskey
Third Advisor
Yujin Kim
Fourth Advisor
Anbu Thamizhchelvan
Fifth Advisor
Brian DeHaven, Ph.D.
Abstract
Multiple myeloma (MM) is a plasma cell disorder accounting for about 10% of hematological malignancies worldwide. Median survival remains at roughly five years post-diagnosis, and long-term remissions are rare. The disease so far has been hard to cure is due to protection provided by soluble factors within the innermost cellular layer of bone, known as the bone marrow microenvironment. Lab simulations are generally inadequate in drug development; therefore, developing a realistic in vitro model of the bone marrow microenvironment is paramount to better understanding the disease. Bortezomib, a dipeptide boronic acid, serves as a chemotherapeutic inhibitor of the 26S proteasome and is used as part of the initial therapies for transplant-eligible patients of MM. Apoptotic activation is a well-supported hypothesis in myeloma cells treated with bortezomib. This study aims to evaluate the time and dosage-dependent inhibitory effects of bortezomib on MM cell growth in a traditional two-dimensional (2D), a transwell 2D, and a bio-printed three-dimensional (3D) cell culture model. Our results indicated that cell death occurred in RPMI-8226 cells grown in the traditional well plate over a period of 72 hours. Markers of apoptosis were successfully isolated and blotted from myeloma cells cultured in traditional well plates and transwell plates. Flow cytometric analysis indicated a decrease in viability and an increase in apoptotic signaling with increasing treatment concentrations. Finally, a 3D gelatin-based bioprinted construct composed of only myeloma cells was generated to determine the cytotoxic effects of 2.5 nM of bortezomib. Results showed a 45% decrease in viability within 48 hours. In conclusion, we generated in-vitro models to test an anti-myeloma agent.
Recommended Citation
Ndubuisi, Chimaobi, "Development of Cell Culture Models for Multiple Myeloma Drug Discovery: Comparison of Traditional 2D, Transwell 2D and 3D-Bioprinted Models" (2024). PCOM Biomedical Studies Student Scholarship. 233.
https://digitalcommons.pcom.edu/biomed/233