Orientational Culture and Evaluation of Endothelial Cells from Human Bone Marrow Stromal Cells in Vitro

Abstract: Human bone marrow stromal cells (BMSCs) were separated by gradient centrifugation on Ficoll (density 1.077 g/ml) from human bone marrow (HBM) in vitro， orientational cultured and amplified endothelial cells (ECs) in DMEM (high glucose) with 20% fetal bovine serum (FBS)， GM-CSF (100 u/ml)， VEGF(10 ng/ml)， FGF(5 ng/ml)， L-glutamine (2 mmol/L)， hepalean (90 u/ml) and antibiotics for about 21-28 days. The property of these ECs was evaluated by morphology with light microscopy and transmission electron microscopy (TEM)， and immunohistochemistry with factor VIII related antigen. After orientational cultured for endothelial cells， 5.0×105 of BMSCs in the primary culture was increased to 6.0×109 of ECs， or to increase 1.2×104 times after 8 generations incubated. More than 70%-80% of the cells from BMSCs cultured after 21-28 days were positive stain for factor VIII related antigen by immunohistochemistry assay. The typical "cobblestone" of these cells presented was observed by light microscopy. The Weible-palade corpuscle， which form is typical morphology of ECs， was also observed by TEM in the cytoplasm. The results showed that BMSCs from HBM had the capability in orientational culture for ECs， which after amplification in vitro might be another source of the seed cells in the cardiovascular tissue-engineering

CSTR: 32200.14.cjcb.2004.06.0019