Hailey Cunningham
Research Mentor(s): Todd Herron
Department or Program: UROP-Cardiovascular Regeneration Core Lab
Authors: Hailey Cunningham, Prakaimuk Saraithong, Devika Baddhan, Sama Salih, Todd J. Herron
Session: Session 2: 1:00pm-1:50pm
Poster: 11
Abstract
Human stem cell-derived heart muscle cells hold promise to regenerate failing hearts; however, current in vitro biomanufacturing processes do not provide enough healthy cells for regeneration therapies. New biomanufacturing processes are required to improve the efficiency and yield of human stem cell-derived cardiomyocytes (heart muscle cells). This summer, we conducted experiments to validate a highly innovative Artificial Intelligence (AI) guided laser cell processing method for the large-scale manufacture of human stem cell-derived cardiomyocytes (heart muscle cells). We used two separate human-induced pluripotent stem cell (hiPSC) lines to test the efficiency of AI laser cell processing for in vitro cardiomyocyte production. First, each stem cell line was plated in separate 6-well plates specially manufactured for use in the Kataoka Cell Processing Device (Kataoka Corporation, CPD-017), and cardiac-directed differentiation was executed using an established small molecule-based protocol to generate cardiomyocytes. The CPD-017 machine contains a small single-slot incubator (37°C and 5.0% CO2) for environmental control, and the device software utilizes AI for image-based discrimination between cardiomyocytes (wanted cells) and non-cardiomyocytes (unwanted cells). First, we scan the plate and observe the “wanted cell area” to start the laser purification process. This area reports the percentage of cardiomyocytes (preferably 10% or more) relative to the percentage of total cells in each well. In longitudinal AI detection of cardiomyocytes, this percentage routinely increased from day 5 to day 8. After day 8 of differentiation, we processed the plates, which means we programmed the laser to kill all the unwanted cells. The health of the AI-laser processed cardiomyocytes (wanted cells) was assessed using Annexin V-Red reagent, which binds to and labels dead/dying cells Red. After laser processing, cardiomyocytes were negative for Annexin V-Red reagent, and dead/dying cells were restricted to the laser-irradiated regions. The new AI-laser processing method yields 20,000,000 cardiomyocytes per 6-well plate (four times more than the old method). Our validation efforts indicate that cardiomyocyte-AI laser processing yields a high number of healthy cardiomyocytes that can now meet the demand for the development of heart regeneration therapies.
