Tailored environments for directed mesenchymal stromal cell proliferation and differentiation using decellularized...

Area of Science:

• Biomaterials Science
• Cell Biology
• Tissue Engineering

Background:

• Mesenchymal stromal cells (MSCs) are vital for regenerative medicine, but controlling their ex vivo behavior is crucial.
• Decellularised extracellular matrix (dECM) from MSCs shows promise for improving MSC culture.
• Previous studies often used stiff substrates, potentially biasing MSC differentiation.

Purpose of the Study:

• To investigate how substrate modulus influences dECM properties and MSC behavior.
• To determine if tailored dECM/substrate combinations can optimize MSC proliferation and differentiation.
• To explore the mechanistic role of YAP1 in MSC response to substrate stiffness.

Main Methods:

• Produced dECM on polyacrylamide hydrogels with varying moduli (4, 10, 40 kPa) and a gradient.
• Cultured MSCs on these dECM-coated substrates.
• Characterized dECM composition using mass spectrometry and fluorescence spectroscopy.
• Assessed MSC proliferation, osteogenic differentiation, and YAP1 nuclear translocation.

Main Results:

• dECM composition varied with substrate modulus.
• Softer substrates (4 kPa) with dECM significantly increased MSC proliferation (1.6-fold vs. glass).
• Higher modulus substrates (40 kPa, glass) with dECM enhanced osteogenic differentiation.
• YAP1 nuclear translocation occurred on substrates ≥10 kPa, correlating with osteogenesis.

Conclusions:

• Integrating dECM technology with substrate modulus control allows for tailored MSC culture environments.
• This approach can optimize MSC proliferation and lineage-specific differentiation for therapeutic applications.
• Results advance the development of advanced tissue engineering scaffolds and MSC-based therapies.