COLLECT EXTRACELLULAR MATRIX FROM IN VITRO FIBROBLAST

Abstract

Extracellular matrices (ECM) have been reported to enhance cell attachment and proliferation as well as to create stem cell niches invitro. We harvested ECM from human fibroblasts for a number of researches, including tissue engineering. Fibroblasts were isolated from human foreskins, cultured in DMEM/F12 containing 10% FBS and identified by Trichrome staining and immunohistochemistry for Vimentin. Then, fibroblasts were stimulated to synthetize ECM in medium supplemented 0.05% ascorbic acid. Cell constituents were removed by using Triton X-100, NH4OH and DNAse. ECM proteins were evaluated by PAS staining. Results showed that collagen is present in ECM.