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DOI: 10.1055/s-2007-967541
Cryopreserved human bone marrow progenitor cells as a potential cell source for cardiovascular tissue engineering
Objective: The suitability of human bone marrow stem cells as an autologous cell source for heart valve tissue engineering has been demonstrated before. For establishment of clinical-grade cell banks cryopreservation will be necessary to store cells until the patient is ready to receive the implantation, but the impact of freezing, thawing, and prolonged storage on the functional properties of BPMC is unclear. We evaluated the suitability of cryopreserved human bone marrow progenitor cells (BMPC) for fabrication of tissue engineered cardiovascular structures.
Methods: Human BMPC from young donors were isolated and characterized before they were cryopreserved for 8–10 weeks. After thawing the cells underwent the same profiling studies. Pre-coated poly-4-hydroxy-butyrate (P4HB) patches were then seeded with cryopreserved BMPC, incubated for various time periods, and examined by light microscopy, immunohistology, and electron microscopy (TEM). Functional properties such as intracellular Ca2+ fluxes in response to pharmacologic stimulation were measured by immunofluorescence imaging.
Results: Cryopreserved BMPC attached to P4HB patches similar to fresh BPMC, and migrated efficiently into the polymer material. Immunohistology and TEM demonstrated morphology cell elements typical of viable cells as well as the expression of extracellular matrix proteins including collagen and fibronectin. Ca2+-flux in response to histamin stimulation showed a pattern comparable to that of fresh BPMC, demonstrating the viability and functional capacity of cryopreserved cells.
Conclusion: For generation of tissue engineered constructs, cryopreserved human BMPC are as effective as freshly isolated cells. Cryopreserved BPMC seeded onto polymeric scaffolds have viability and functional properties suitable for potential cardiovascular applications.