Serum deprivation improves seeding and repopulation of acellular matrices with valvular interstitial cells.
Cell-extracted valvular tissues (acellular scaffolds, or aScaffolds) offer unique advantages over synthetic polymers for cardiac valve engineering applications in that they retain extracellular matrix molecules to support cellular ingrowth. The extracellular matrix is important in directing many cellular pathways, such as adhesion, proliferation, migration, differentiation, and survival. However, repopulating this type of scaffold often requires high seeding densities or recurrent cell delivery. The optimization of valvular interstitial cell (VIC) seeding onto aScaffolds is reported herein. VICs (the most prevalent cell type in valve leaflets) have maximal growth in 15-20% serum concentrations on tissue-culture polystyrene. Interestingly, after VIC seeding onto aScaffolds, a reduction of serum content, from 15% serum to 5% or less, was found to increase significantly the number of adherent cells, as well as induce transfer of VICs from a tissue-culture polystyrene surface to the aScaffold. aScaffolds seeded and cultured with periods of reduced serum levels were shown to support and enhance VIC viability and attachment, as well as accelerate VIC migration into the aScaffold, leading to a uniformly repopulated valve leaflet construct after 4 weeks of static culture.
Cushing
Melinda C
MC
Department of Chemical and Biological Engineering, ECCH 111, UCB 424, University of Colorado, Boulder, CO 80309-0424, USA.
Jaeggli
Michael P
MP
Masters
Kristyn S
KS
Leinwand
Leslie A
LA
Anseth
Kristi S
KS
D000818:Animals; D001021:Aortic Valve / Q000166:cytology*; D001672:Biocompatible Materials; D002448:Cell Adhesion; D002454:Cell Differentiation; D002465:Cell Movement; D049109:Cell Proliferation; D002470:Cell Survival; D002478:Cells, Cultured; D016895:Culture Media, Serum-Free / Q000494:pharmacology; D004247:DNA / Q000737:chemistry; D005109:Extracellular Matrix / Q000378:metabolism; D006351:Heart Valves / Q000166:cytology / Q000378:metabolism*; D018613:Microscopy, Confocal; D013499:Surface Properties; D013552:Swine; D013778:Tetrazolium Salts / Q000494:pharmacology; D013844:Thiazoles / Q000494:pharmacology; D013997:Time Factors; D023822:Tissue Engineering / Q000379:methods*
D001672:Biocompatible Materials; D016895:Culture Media, Serum-Free; D013778:Tetrazolium Salts; D013844:Thiazoles; D004247:DNA; C022616:thiazolyl blue