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Cryopreservation of hMSCs seeded silk nanofibers based tissue engineered constructs
Akalabya Bissoyi1, K Pramanik1, Niladri Nath Panda1
1Department of Biotechnology and Medical Engineering, National Institute of Technology, Rourkela, India.
Cryobiology
|April 25, 2014
Summary
This study developed an effective freezing medium for tissue engineering constructs (TECs) using natural osmolytes like trehalose and ectoin. This method ensures long-term preservation of TECs, maintaining cell viability and differentiation potential.
Area of Science:
- Biomaterials Science
- Regenerative Medicine
- Cryobiology
Background:
- Long-term cryopreservation of tissue engineering constructs (TECs) is crucial for off-the-shelf availability in medical applications.
- Conventional cryopreservation methods often face challenges in maintaining cell viability and function post-thaw.
Purpose of the Study:
- To evaluate the efficacy of natural osmolytes (trehalose, ectoin) with or without dimethyl sulfoxide (Me2SO) for cryopreserving TECs.
- To formulate an optimized freezing medium for long-term TEC preservation.
Main Methods:
- Umbilical cord-derived mesenchymal stem cells (MSCs) were cultured on silk fibroin scaffolds to create TECs.
- TECs were cryopreserved using nine different freezing solutions, including combinations of natural osmolytes and Me2SO.
- Post-thaw analysis included cell viability (PI staining), proliferation (MTT assay), microstructure (SEM), membrane integrity (confocal microscopy), and osteogenic differentiation (ALP, RT-PCR, histology).
Main Results:
- A freezing medium containing trehalose (40mM), ectoin (40mM), catalase (100μg), and Me2SO (2.5%) proved most effective.
- The optimized medium maintained TEC integrity, MSC viability, and osteogenic differentiation potential post-thaw.
- Cryopreserved TECs exhibited mechanical integrity comparable to non-cryopreserved controls and superior performance to conventional Me2SO-based media.
Conclusions:
- An efficient freezing medium utilizing natural osmolytes has been formulated for long-term cryopreservation of TECs.
- This formulation supports the preservation of TEC integrity, MSC viability, and differentiation capacity.
- The developed method offers a promising approach for the clinical application of tissue engineering constructs.
