ABSTRACT
Hepatic progenitor cells (HPCs) have regenerative properties that could aid the development of treatments for severe liver disease. To study how pressure influences HPC fate, a hydrostatic pressure-controlled cell culture chamber was developed. The design incorporates custom LabView scripting for enhanced pressure regulation and data acquisition. Pressure can be controlled within ±0.2mmHg. Continuous airflow permits gas exchange, and CO2 is maintained at 5%±0.2%. Applied pressures range from 5 to 20 mmHg, reflecting interstitial pressure conditions in healthy and diseased livers, respectively. Bipotential Murine Oval Liver (BMOL) cells, an HPC-like cell line, were cultured in the chamber to test for maintenance of cell viability, adequate CO2 regulation, and maintenance of adequate media volume over 24 hours. Cultured cells were exposed to 5 or 19 mmHg. After 24 hours, media pH was measured, viable cells were counted (Trypan Blue, n=3), and plates were weighed to assess fluid loss. The number of live cells cultured under pressure vs. control conditions was not statistically different (p>.05). The pH remained constant at 7.0 for all conditions, suggesting adequate gas exchange. Evaporation of media was minimal at 3.97%. Results indicate that the pressure chamber provides appropriate environmental conditions for future studies on HPC pressure sensitivity.
