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Slow cooling cryopreservation of cell-microcarrier constructs
Evi Lippens1, Maria Cornelissen
1Department of Basic Medical Sciences, Ghent University, Ghent, Belgium.
Cells, Tissues, Organs
|April 22, 2010
Summary
Cryopreserving cell-loaded bone tissue constructs using dimethyl sulfoxide (Me2SO) with or without ascorbic acid (AA) shows promise for faster availability. This method may reduce the need for extensive cell culture periods before implantation in bone defect repair.
Area of Science:
- Biomaterials Science
- Regenerative Medicine
- Cell Biology
Background:
- Tissue-engineered constructs are crucial for treating complex bone defects.
- Prolonged cell culture periods limit the immediate availability of these constructs.
- Cryopreservation offers a potential solution to overcome culture time limitations.
Purpose of the Study:
- To investigate the feasibility of cryopreserving cell-loaded tissue engineered constructs.
- To evaluate the impact of different cryomedium formulations on cell viability and construct recovery.
- To assess the potential for reducing pre-implantation culture time.
Main Methods:
- Goat bone marrow-derived mesenchymal stem cells (BMSC) and MC3T3-E1 cells were cultured on microcarriers.
- Constructs were cryopreserved using slow cooling to -80°C and stored in liquid nitrogen.
- Four cryomedia formulations were tested, including varying concentrations of dimethyl sulfoxide (Me2SO), ascorbic acid (AA), hydroxyethyl starch, and sucrose.
- Cell viability was assessed post-thaw using live/dead staining and MTS assay.
Main Results:
- Immediately after thawing, cell viability was significantly reduced across all tested cryomedia.
- No significant differences in viability were found between the different cryomedia formulations.
- A trend suggested higher cell survival and faster recolonization with Me2SO-based cryomedia, with or without AA.
- Recolonization periods were approximately 3 days for BMSC and 3.6-3.8 days for MC3T3-E1 constructs in Me2SO-based media.
Conclusions:
- Cryopreservation of cell-loaded constructs is feasible, though initial viability is reduced.
- Me2SO-based cryomedia, with or without AA, appear most promising for cell survival and construct recovery.
- This approach could potentially shorten the time required for tissue-engineered construct preparation, facilitating faster clinical application.
