Share this page on LinkedIn
Share This Page on Google+
Share This Page on Twitter
tell someone about this page print this page
You are here: Contents > 2010 > Volume 19 Number 6 November 2010 > MOLECULAR BIOLOGY > Functional Characterization of Fibronectin-Separated Valve Interstitial Cell Subpopulations in Three-Dimensional Culture

Functional Characterization of Fibronectin-Separated Valve Interstitial Cell Subpopulations in Three-Dimensional Culture

Elizabeth H. Stephens, Joshua L. Carroll, Allison D. Post, Joyce J. Kuo, K. Jane Grande-Allen 

Department of Bioengineering, Rice University, Houston, Texas, USA

Background and aim of the study: Myxomatous mitral valves (MVs) contain elevated proportions of myofibroblasts, a valve interstitial cell (VIC) subpopulation that may be important in disease pathogenesis. A novel technique was recently developed for the isolation of VIC myofibroblasts using time-dependent adhesion to fibronectin (FN). Cells that adhere rapidly to FN (‘FAST’) demonstrate myofibroblast cell phenotype markers, in contrast to cells that fail to adhere after a longer time (‘SLOW’). The study aim was to characterize the functionality of these subpopulations using three-dimensional (3D) collagen constructs.
Methods: The VICs were harvested from porcine mitral valve posterior leaflets. FAST and SLOW subpopulations, as well as unseparated VIC populations grown on FN and tissue culture plastic (TCP) (UNSEP FN, UNSEP TCP), were seeded within 3D collagen gels and cultured for three weeks. Collagen gel contraction was assessed throughout the culture duration; the mechanical properties of the resultant collagen constructs were assessed using uniaxial tensile testing.

Results: FAST cells demonstrated a greater contraction of collagen gels compared to SLOW cells, particularly after 10 days (p <0.05). Interestingly, the collagen gel contraction by both FN-separated VIC subpopulations (FAST and SLOW) was greater than for gels seeded with UNSEP TCP VICs (p <0.05). Further, the contraction of UNSEP FN gels was greater than UNSEP TCP throughout the culture duration (p ?0.002), suggesting that the subculture of VICs on FN potentiated these phenotypic changes. Finally, the collagen constructs seeded with FAST cells were stiffer than those seeded with SLOW, followed by UNSEP TCP (p <0.001). The same pattern was found for failure stress (p = 0.006).
Conclusion: Time-dependent adhesion to FN produced a VIC subpopulation (FAST), the function of which in 3D culture was consistent with that of myofibroblasts; FN exposure alone also caused VICs to function similarly to myofibroblasts. This novel isolation method may prove valuable in future studies of myofibroblasts in valve disease.

The Journal of Heart Valve Disease 2010;19:759-765

Functional Characterization of Fibronectin-Separated Valve Interstitial Cell Subpopulations in Three-Dimensional Culture

Click the above hyperlink to view the article, right click (Ctrl click on a Mac) to open in a new browser window or tab.

Purchase this Article

Please click the button below to purchase this article. Single article purchases are provided at $50.00 per article. Upon clicking the button below, single article user account subscription details are requested and, upon successful payment, a single article user account is created. Single articles are availble in your account for seven days after purchase.