Peristaltic Pump to Automate Media Flow for Tissue-Engineered Muscle Repair (TEMR) Construct; How do Ethical Concerns Regarding CRISPR/Cas9 and Stem Cell Therapy Affect the Research for Volumetric Muscle Loss (VML)? 325 views

Volumetric Muscle Loss (VML) injuries result in functional loss in the muscle where more tissue is lost than the body can regenerate or repair. The Christ Lab at UVA has developed a Tissue Engineered Muscle Repair construct (TEMR) to help with treatment of VML injuries. However, contamination can easily be introduced during the TEMR biomanufacturing process following media changes, since the bioreactor used to incubate the TEMRs must be opened every forty-eight hours. To address this issue, we developed a perfusion system using two peristaltic pumps derived from an open-source design to change media in the bioreactor automatically while maintaining a closed system. We adapted a rotor to provide more torque than was inherent in the original design, which relied on static friction, increasing the range of possible flow rates. After calibrating motor speed to flow rate, we tested three parameters that impact media perfusion into and out of the bioreactor: 1) flow rate, 2) volume pumped in/out at once, and 3) pause time between pumping stages. Using a dye diffusion technique where dyed water simulated fresh media, we found that the peristaltic pumps were able to create a homogenous distribution of dye throughout the bioreactor during a half-media change, and that pause time does not significantly affect the concentration of dye in the wastewater. Furthermore, the results showed a flow rate of 1.5 mL/min with 3 mL of media exchange resulting in the lowest dye concentration in waste. These findings indicate that peristaltic pumps are a valid and efficient way to perfuse fresh media and discard depleted media.

BS (Bachelor of Science)

Volumetric Muscle Loss ; Peristaltic Pump ; Automation ; Ethical Concerns ; CRISPR/Cas9; Stem Cell Therapy

School of Engineering and Applied Science Bachelor of Science in Biomedical Engineering Technical Advisor: Megan Lobdell, George Christ STS Advisor: Pedro Francisco Technical Team Members: Abigail Boyette, Eirian Crocker

English

2024-05-09