Amplification of mesenchymal stemcells from human bone marrow and ori-entation to induce MSCs differentiating into endothelial cells in vitro
- VernacularTitle:人骨髓间质干细胞体外扩增和向内皮细胞定向诱导分化的研究
- Author:
Bin FENG
;
Ying-Long LIU
;
Kai FENG
;
Ru GONG
;
Hu CHEN
;
- Publication Type:Journal Article
- Keywords:
Bone marrow;
Mesenchymal stemcells;
Endothelial cells
- From:
Chinese Journal of Pathophysiology
2000;0(08):-
- CountryChina
- Language:Chinese
-
Abstract:
AI M:Our purpose wastoinduce MSCs differentiatinginto endothelial cells (EC)invitroandto providetheseed cells for study of cardiovascular tissue -engineering.METHODS :MSCs were separated by gradient centrifugation on Percoll(density 1 073 g/L) fromhuman bone marrow(HBM) ,and incubated for purification and amplification in DMEM(lowglucose)with 10 %fetal bovine serum(FBS) .Then,the MSCs were incubated for orientation differentiated into ECin DMEM(high glu-cose) with20 %FBS,VEGF(10?g/L) ,bFGF(5?g/L) ,L-glutamine (2 mmol/L) ,penicillin (1?105U/L) and streptomycin(100 mg/L) for about 14 -21 days andtheir phenotypic characteristics were analyzed byflowcytometry.Afterwards ,the differenti-ating cells were evaluated by histology andimmunohistochemistry.RESULTS :The quantity of MSCs was increasedfrom5?0?105inthe primary culture to 8?0?1012,or toincrease to 1?6?107times after 15 generations of incubation.The purity of MSCs wasabove 95 %and 98 %homogeneous at passages 2 and 3 ,respectively.About 80 %-90 %of the differentiating cellsfrom MSCs af-ter 14 -21 days were positively stainedfor Ⅷfactor (vWF) related antigen by immunohistochemistry assay,and Weible -paladecorpuscle was also observed bytransmission electron microscopyinthe cytoplasm.CONCLUSION:MSCs from HBMhave the ca-pability of differentiationinto ECsin vitro,which may be a potential source of seed cellsforfabrication of tissue -engineering heartvalve ,particularlyin children with congenital heart disease .
