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Vitreous cryopreservation of nanofibrous tissue-engineered constructs generated using mesenchymal stromal cells.
- Source :
-
Tissue engineering. Part C, Methods [Tissue Eng Part C Methods] 2009 Mar; Vol. 15 (1), pp. 105-14. - Publication Year :
- 2009
-
Abstract
- Development of an effective preservation strategy to fulfill off-the-shelf availability of tissue-engineered constructs (TECs) is demanded for realizing their clinical potential. In this study, the feasibility of vitrification, ice-free cryopreservation, for precultured ready-to-use TECs was evaluated. To prepare the TECs, bone marrow-derived porcine mesenchymal stromal cells (MSCs) were seeded in polycaprolactone-gelatin nanofibrous scaffolds and cultured for 3 weeks before vitrification treatment. The vitrification strategy developed, which involved exposure of the TECs to low concentrations of cryoprotectants followed by a vitrification solution and sterile packaging in a pouch with its subsequent immersion directly into liquid nitrogen, was accomplished within 11min. Stepwise removal of cryoprotectants, after warming in a 38 degrees C water bath, enabled rapid restoration of the TECs. Vitrification did not impair microstructure of the scaffold or cell viability. No significant differences were found between the vitrified and control TECs in cellular metabolic activity and proliferation on matched days and in the trends during 5 weeks of continuous culture postvitrification. Osteogenic differentiation ability in vitrified and control groups was similar. In conclusion, we have developed a time- and cost-efficient cryopreservation method that maintains integrity of the TECs while preserving MSCs viability and metabolic activity, and their ability to differentiate.
- Subjects :
- Alkaline Phosphatase metabolism
Animals
Anthraquinones
Calcium metabolism
Cell Proliferation
Cell Shape
Cell Survival
Cells, Cultured
Collagen Type I metabolism
Mesenchymal Stem Cells enzymology
Nanostructures ultrastructure
Osteogenesis
Stromal Cells enzymology
Surface Properties
Sus scrofa
Tissue Scaffolds
Cryopreservation methods
Mesenchymal Stem Cells cytology
Nanostructures chemistry
Stromal Cells cytology
Tissue Engineering
Subjects
Details
- Language :
- English
- ISSN :
- 1937-3384
- Volume :
- 15
- Issue :
- 1
- Database :
- MEDLINE
- Journal :
- Tissue engineering. Part C, Methods
- Publication Type :
- Academic Journal
- Accession number :
- 19196127
- Full Text :
- https://doi.org/10.1089/ten.tec.2008.0237