Preparation and evaluation of extracellular matrix scaffold of human adipose tissue

Abstract

25 g normal adipose tissue was cut into pieces. Then repeated freeze-thaw,enzymatic digestion,organic solvent extraction and vacuum freeze-drying were performed.Adult adipose tissue extracellular tissue matrix was obtained. The traits of extracellular matrix scaffold were observed.HE staining, Masson staining and DAPI fluorescence staining were used to test the effectiveness of the decellularization.Immunohistochemistry was used to detect the reservations of extracellular matrix (Ⅳ collagen, laminin).Scanning electron microscopy was introduced to observe the ultrastructure of extracellular matrix scaffold, and universal mechanical testing machine was used to measure the mechanical properties of the scaffolds. Enzyme digestion method was used to extract human adipose-derived stem cells (hADSCs),and then the 3rd passage hADSCs were cocultured with extracellular matrix scaffold.CCK8 was introduced to assay cell proliferation activity,and scanning electron microscopy was used to observe cellular adhesion. The cells on the scaffold were induced to adipocytes and observed by freezing section and Oil Red O staining after 14 days. The extracellular matrix scaffold of adipose tissue was porous sponges architecture. The cells in tissue were relatively removed. The collagen and laminin were preserved relatively, and the mechanical properties did not decline too much. The cell proliferation, adhesion and differentiation on the scaffold was very well. The extracellular matrix scaffold of adipose tissue, prepared through the methods of physical, chemical, enzymatic digestion and vacuum freeze-drying method, keeps the main ingredients of extracellular matrix and presents a well cellular compatibility, therefore it should be an ideal biologic scaffold for adipose tissue engineering.