Functional Profiling of Chondrogenically Induced Multipotent Stromal Cell Aggregates Reveals Transcriptomic and...

Area of Science:

• Biomedical Engineering
• Cell Biology
• Regenerative Medicine

Background:

• Multipotent stromal cells (MSCs) are promising for bone and cartilage repair.
• Functional variability in MSCs hinders clinical use, necessitating better assessment of chondrogenic capacity.
• Current methods for evaluating MSC chondrogenesis are insufficient for manufacturing.

Purpose of the Study:

• To develop a high-throughput, nondestructive method for assessing MSC chondrogenic potential.
• To identify specific morphological features of MSC aggregates that correlate with chondrogenesis.
• To enable selection of MSC lines with optimal chondrogenic capacity for clinical applications.

Main Methods:

• Chondrogenic induction of MSC aggregates from eight human cell lines across multiple passages.
• Dynamic monitoring of aggregate morphology using functionally relevant profiling.
• Quantitative functional assays to assess long-term chondrogenesis.
• Correlation analysis between morphological features and chondrogenic outcomes.

Main Results:

• MSC aggregate morphology exhibited unique, cell line- and passage-dependent dynamics.
• Specific morphological features (spheroid area, radius, minimum feret diameter, minor axis length) correlated with chondrogenic synthetic activity by day 4.
• Morphological profiling predicted long-term chondrogenesis, independent of gene expression changes at early time points.

Conclusions:

• Dynamic morphological profiling is a sensitive, early indicator of MSC chondrogenic capacity.
• This approach can guide the selection of MSCs for improved cartilage tissue engineering.
• The method offers a valuable tool for optimizing MSC manufacturing for regenerative medicine products.