Sequential use of human-derived medium supplements favours cardiovascular tissue engineering.
Bottom Line: As a prove of principle, 3D tissue culture and tensile testing was performed for two patients, to determine tissue strength.Tensile testing showed increased strength for tissues cultured in HS when compared to PL.This effect was more pronounced if cells were sequentially cultured in PL, followed by tissue culture in HS.
Affiliation: Department of Cardio-Thoracic Surgery, University Medical Center, Utrecht, The Netherlands. firstname.lastname@example.orgShow MeSH
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Mentions: Figure 4A shows zymographic analysis of gelatinolytic activity corrected for their presence in non-conditioned media, and displays release of active and inactive forms of MMP2 (72 and 64 kD) and active MMP9 (82 kD) by the cells in both PL- and HS-conditioned cells, but not of inactive MMP9 (92 kD). When expression in PL was compared to HS, no significant differences were found for inactive forms of MMP2 and MMP9 and active MMP9 [averages PL: 27718 ± 6803, 0 ± 4479 and 4306 ± 1701 and HS: 23299 ± 5299, 91 ± 3151 and 6105 ± 1640 arbitrary units (a.u.) respectively], but quantification of active MMP2 revealed a significant twofold increase in PL-cultured cells (PL: 6215 ± 1278 versus HS 3608 ± 1375 a.u.). On the collagen-substituted gels (Fig. 4B), we did not find significant differences in MMP-activity between cells cultured in PL or HS. Taken together, zymography analysis revealed a reduced matrix-degradation capacity through a significant decrease of active MMP2 levels when cells are cultured in HS-supplemented medium.
Affiliation: Department of Cardio-Thoracic Surgery, University Medical Center, Utrecht, The Netherlands. email@example.com