Differential Phase Contrast Imaging to Predict MSC Immune Function.

Area of Science:

• Cell Biology
• Immunology
• Biotechnology

Background:

• Mesenchymal stromal cells (MSCs) possess immunomodulatory properties beneficial for treating autoimmune and inflammatory diseases.
• Clinical translation of MSCs is limited by culture heterogeneity and the absence of reliable potency assays.
• Indoleamine-2,3-dioxygenase (IDO) activity is a key measure of MSC immunosuppressive function.

Purpose of the Study:

• To investigate single-cell morphological imaging as a non-invasive method to estimate MSC indoleamine-2,3-dioxygenase (IDO) activity.
• To develop machine learning models for predicting IDO activity from live MSCs during biomanufacturing.
• To extend imaging characterization for assessing MSC potency based on IDO activity and T-cell immune suppression.

Main Methods:

• Utilized fluorescence and label-free quantitative differential phase contrast (qDPC) imaging to capture live MSC morphology.
• Extracted morphological features from single MSCs using quantitative imaging techniques.
• Applied machine learning (ML) regression models to predict single-cell IDO protein and enzyme activity.

Main Results:

• Successfully extracted morphological features from live MSCs using qDPC imaging.
• Developed ML models capable of predicting single-cell IDO activity from morphological data.
• Demonstrated that qDPC imaging can estimate MSC potency, correlating with IDO activity and T-cell suppression.

Conclusions:

• Single-cell morphological imaging offers a scalable, non-destructive approach to monitor MSC immunomodulatory capacity.
• This method provides a foundation for developing robust quality control strategies in MSC manufacturing.
• The findings support the advancement of MSC-based therapies for inflammatory and autoimmune conditions.