Tracking protein function with sodium multi quantum spectroscopy in a 3D-tissue culture based on microcavity arrays

Area of Science:

• Biochemistry
• Cell Biology
• Medical Imaging

Background:

• The Na-/K-ATPase enzyme is crucial for cellular function.
• Monitoring enzyme activity in real-time within a controlled environment is challenging.
• Magnetic Resonance (MR) techniques offer non-invasive monitoring capabilities.

Purpose of the Study:

• To investigate the effects of strophanthin-induced Na-/K-ATPase inhibition in liver cells.
• To demonstrate the utility of a novel MR-compatible bioreactor for cell culture studies.
• To validate the use of sodium multi-quantum (MQ) spectroscopy for assessing enzyme activity.

Main Methods:

• Utilized a microcavity array for high-density, three-dimensional liver cell culture within an MR-compatible bioreactor.
• Employed direct contrast-enhanced (DCE) MRI to confirm homogenous substance distribution.
• Applied a time proportional phase increment (TQTPPI) pulse sequence for simultaneous detection of single quantum (SQ) and triple quantum (TQ) MR signals.

Main Results:

• The MR-compatible bioreactor allowed precise control over physiological parameters (temperature, gas, fluid flow).
• Sodium TQ/SQ MR signals accurately reflected the time course of Na-/K-ATPase inhibition.
• Homogenous distribution of biochemical substances within the bioreactor was confirmed by DCE MRI.

Conclusions:

• The developed MR-compatible bioreactor is a valuable tool for studying cellular processes in real-time.
• Sodium multi-quantum spectroscopy, particularly with TQTPPI, provides accurate insights into Na-/K-ATPase activity.
• This approach enables detailed observation of enzyme inhibition dynamics in a controlled cellular environment.