Limits...
Sequential use of human-derived medium supplements favours cardiovascular tissue engineering.

Riem Vis PW, Sluijter JP, Soekhradj-Soechit RS, van Herwerden LA, Kluin J, Bouten CV - J. Cell. Mol. Med. (2012)

Bottom Line: 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.These data suggest that sequential use of PL and HS as substitutes for FBS in culture medium for cardiovascular tissue engineering results in improved cell proliferation and tissue mechanical properties, as compared to use of PL or HS apart.

View Article: PubMed Central - PubMed

Affiliation: Department of Cardio-Thoracic Surgery, University Medical Center, Utrecht, The Netherlands. priemvis@umcutrecht.nl

Show MeSH

Related in: MedlinePlus

(A) Quantification of MMP2 and MMP9 expression by cells in culture (mean ± S.D.), corrected for unconditioned medium (UCM). Cells cultured in both media produce and activate MMP2, whereas MMP9 did not show a difference compared to the unconditioned control. Activated MMP2 expression was significantly lower in the cells cultured with HS (**P < 0.01). The inlay shows an example of expression patterns by one patient and the size of the signals that were quantified. A.U.: arbitrary units as a measure for number of counted pixels in a pre-determined surface. (B) Quantification of collagenases by cells in culture (mean ± S.D.). Expression patterns are presented in the inlay. We did not find significant differences in collagenase expression between cells cultured in PL or HS. Up: MMPs expressed at marks A and B; Low: MMPs expressed at marks C and D. Active forms (Act, B and D), are slightly smaller in size when compared to inactive forms (Inact, A and C) of these MMPs.
© Copyright Policy
Related In: Results  -  Collection


getmorefigures.php?uid=PMC3822844&req=5

fig04: (A) Quantification of MMP2 and MMP9 expression by cells in culture (mean ± S.D.), corrected for unconditioned medium (UCM). Cells cultured in both media produce and activate MMP2, whereas MMP9 did not show a difference compared to the unconditioned control. Activated MMP2 expression was significantly lower in the cells cultured with HS (**P < 0.01). The inlay shows an example of expression patterns by one patient and the size of the signals that were quantified. A.U.: arbitrary units as a measure for number of counted pixels in a pre-determined surface. (B) Quantification of collagenases by cells in culture (mean ± S.D.). Expression patterns are presented in the inlay. We did not find significant differences in collagenase expression between cells cultured in PL or HS. Up: MMPs expressed at marks A and B; Low: MMPs expressed at marks C and D. Active forms (Act, B and D), are slightly smaller in size when compared to inactive forms (Inact, A and C) of these MMPs.

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.


Sequential use of human-derived medium supplements favours cardiovascular tissue engineering.

Riem Vis PW, Sluijter JP, Soekhradj-Soechit RS, van Herwerden LA, Kluin J, Bouten CV - J. Cell. Mol. Med. (2012)

(A) Quantification of MMP2 and MMP9 expression by cells in culture (mean ± S.D.), corrected for unconditioned medium (UCM). Cells cultured in both media produce and activate MMP2, whereas MMP9 did not show a difference compared to the unconditioned control. Activated MMP2 expression was significantly lower in the cells cultured with HS (**P < 0.01). The inlay shows an example of expression patterns by one patient and the size of the signals that were quantified. A.U.: arbitrary units as a measure for number of counted pixels in a pre-determined surface. (B) Quantification of collagenases by cells in culture (mean ± S.D.). Expression patterns are presented in the inlay. We did not find significant differences in collagenase expression between cells cultured in PL or HS. Up: MMPs expressed at marks A and B; Low: MMPs expressed at marks C and D. Active forms (Act, B and D), are slightly smaller in size when compared to inactive forms (Inact, A and C) of these MMPs.
© Copyright Policy
Related In: Results  -  Collection

Show All Figures
getmorefigures.php?uid=PMC3822844&req=5

fig04: (A) Quantification of MMP2 and MMP9 expression by cells in culture (mean ± S.D.), corrected for unconditioned medium (UCM). Cells cultured in both media produce and activate MMP2, whereas MMP9 did not show a difference compared to the unconditioned control. Activated MMP2 expression was significantly lower in the cells cultured with HS (**P < 0.01). The inlay shows an example of expression patterns by one patient and the size of the signals that were quantified. A.U.: arbitrary units as a measure for number of counted pixels in a pre-determined surface. (B) Quantification of collagenases by cells in culture (mean ± S.D.). Expression patterns are presented in the inlay. We did not find significant differences in collagenase expression between cells cultured in PL or HS. Up: MMPs expressed at marks A and B; Low: MMPs expressed at marks C and D. Active forms (Act, B and D), are slightly smaller in size when compared to inactive forms (Inact, A and C) of these MMPs.
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.

Bottom Line: 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.These data suggest that sequential use of PL and HS as substitutes for FBS in culture medium for cardiovascular tissue engineering results in improved cell proliferation and tissue mechanical properties, as compared to use of PL or HS apart.

View Article: PubMed Central - PubMed

Affiliation: Department of Cardio-Thoracic Surgery, University Medical Center, Utrecht, The Netherlands. priemvis@umcutrecht.nl

Show MeSH
Related in: MedlinePlus