A safe and efficient method to retrieve mesenchymal stem cells from three-dimensional fibrin gels

Area of Science:

• Biomaterials Science
• Cell Biology
• Regenerative Medicine

Background:

• Mesenchymal stem cells (MSCs) are promising for therapy but lose multipotency in traditional 2D cultures.
• Three-dimensional (3D) culture systems better mimic in vivo environments for maintaining cell phenotypes.
• Fibrin hydrogels are attractive 3D substrates for cell culture due to their natural origin and biocompatibility.

Purpose of the Study:

• To develop a safe and efficient method for isolating mesenchymal stem cells (MSCs) from 3D fibrin hydrogels.
• To assess the viability and multipotency of MSCs after extraction from fibrin gels.
• To establish a protocol adaptable for retrieving other cell types from 3D fibrin constructs.

Main Methods:

• Utilized fibrin as a 3D culture matrix for MSCs.
• Employed nattokinase, a fibrinolytic enzyme, for cell extraction from fibrin hydrogels.
• Assessed cell viability, proliferation, and multilineage potential post-extraction.

Main Results:

• Developed a novel protocol for isolating MSCs from 3D fibrin gels using nattokinase.
• Demonstrated that nattokinase effectively lyses fibrin while preserving MSC viability.
• Recovered MSCs retained their proliferative capacity and multilineage differentiation potential.

Conclusions:

• Nattokinase provides a safe and efficient means to retrieve viable MSCs from 3D fibrin hydrogels.
• This method supports the therapeutic potential of MSCs cultured in 3D environments.
• The protocol is adaptable for extracting various cell types from fibrin-based constructs for diverse applications.