Primary culture and 3-dimensional scaffold culture of rat gastric smooth muscle cells in vitro

Objective:To study the method of primary culture of SD rat gastric smooth muscle cells by co-collagenase digestion method,and develop a 3-dimensional scaffold culture model for tissue engineering.Methods:Smooth muscle layer of rat was digested by 0.2% 1-type collagenase and 0.05% trypsin.The cultured cells were observed and identified.Expanded cells of P3 were seeded into PLGA constructs with fibrin matrix.Results:The smooth muscle cells were successfully cultured passaged in 2 weeks.The passaged smooth muscle cells grew with typical peak-valley pattern.Im munohistochemicalmethod (S-P method) showed strong expression of α-smooth muscle actin.Seeding of the expanded PC was technically feasible.The 3-dimensional scaffold culture technique provided homogeneous cell distribution without sig nificant cell loss.Conclusion:Our primary cultured SD rat gastric smooth muscle cells with the co-collagenase method can stably grow.The model of 3D PLGA seeded is technically feasible and applies a promising method for creation of new tissue-engineered tissue in vitro.