Vascular smooth muscle cell expansion and functional maintenance in a three-dimensional suspended environment

The scale expansion and functional maintenance of vascular smooth muscle cells (VSMCs) represent one of the challenges in obtaining tissue-engineered blood vessels (TEBV). Microcarrier suspension culture technology combined with surface area advantages and appropriate mechanical stimulation may become an important technical means for its three-dimensional(3D) expansion. Bovine VSMCs were selected as seed cells, and the dynamic culture system based on 3D TableTrix™ microcarrier was employed as the technical platform to optimize the cultivation parameters. Conventional two-dimensional(2D) planar culture was selected for comparative analysis, and the changes in cell performance obtained through the two different expansion techniques were comprehensively evaluated. Expanded VSMCs obtained under optimal technical parameters were used to form TEBV via the cell sheet technique. The efficacy of cells from both expansion techniques in TEBV culture was assessed through tissue section staining and total collagen quantification. Under identical seeding densities of bovine VSMCs, compared with conventional 2D planar culture, 3D dynamic microcarrier culture resulted in a certain degree of expansion difference in total cell count. The difference between the two methods reached 1.43-fold on day 6 and increased to 2.85-fold by day 12. Functional studies of VSMCs obtained via both methods revealed no significant differences in proliferation, migration, phenotype, or collagen secretion ability. TEBV cultured using the cell sheet technique also exhibited no significant differences in tissue architecture or total collagen content. 3D dynamic microcarrier culture supports efficient expansion of bovine VSMCs while maintaining their functional properties.