Biocompatibility of rabbit adipose-derived stem cells with porous polylactic-co-glycolic acid scaffold / 中华实验眼科杂志

ABSTRACT

Background Seed cells and scaffold material are the important aspects of corneal tissue engineering research.Adipose-derived stem cells(ASCs) are becoming the focus of seed cells research because of their wide source,powerful proliferation and differentiation abilities.As biodegradable polymer,polylactic-co-glycolic acid(PLGA) has successfully build multiple tissues and organs.Objective Present study was to ascertain the biological characteristics of the rabbit ASCs and their biocompatibility with PLGA scaffold in vitro and to provide groundwork for further study on the reconstruction of tissue engineered corneal stroma.Methods Adipose cells were isolated from lipoaspirate of New Zealand white rabbit using collagenase Ι.The cells were cultured and passaged.The generation 4 cells were inoculated to culture plate with 6 holes at the density of 3×104/cm2,3×104/cm2,3×106/cm2 respectively and cultivated in ossification inducing medium,lipoblast inducing medium and chondroblast inducing medium to identify the characteristics of the cells.The multilineage differentiated cells were identified by alizarin red staining,oil red O staining and immunoinfluorescene technique.The generation 4 cells were re-suspended with DiO influorescence fluid at the density of 1×107/ml and seeded on PLGA scaffold to fabricate cell-PLGA constructs.Quantitative analysis of cell proliferation on PLGA was detected by Hoechst DNA assay.The attachment and growth of adipose-derived stem cells on the scaffold were observed under the scanning electron microscope(SEM) and confocal microscopy in 1 day,3,7 days after seeding for the evaluation of biocompatibility between cells and PLGA.Results Primarily cultured cells reached 80%-90% confluence after 7-8 days with the fibroblast-like appearance.Adipose-derived stem cells of rabbits differentiated into osteoblast,adipocyte and chondroblast successfully,showing the positive stain for alizarin red staining,oil red O staining and immunoinfluorescene technique respectively.Proliferation of cells on PLGA scaffold went into plateau phase at 7 days after culture.SEM and confocal microscopy revealed the well-attached,spread cells along the scaffold and abundant excellular matrix both on the surface and interior pore of scaffold.Conclusion Cultured rabbit adipose cells have the ability of potential multilineage differentiation and good biocompatibility with PLGA scaffold,which could be used to construction of tissue engineered corneal stroma.