Inducing human bone marrow mesenchymal stem cells to differentiate into endothelial cells with indirect co-culture
- VernacularTitle:非接触共培养法诱导人骨髓间充质干细胞向内皮细胞的分化
- Author:
Jun LU
;
Wei HUANG
;
Xinxin CHEN
- Publication Type:Journal Article
- From:
Chinese Journal of Tissue Engineering Research
2007;0(38):-
- CountryChina
- Language:Chinese
-
Abstract:
BACKGROUND: Recent studies indicate that after indirect co-culture of neonate rat myocardial cells and bone marrow mesenchymal stem cells, bone marrow mesenchymal stem cells can differentiate into myocardial cells successfully. OBJECTIVE: To induce human bone marrow mesenchymal stem cells (hBMSCs) to differentiate into endothelial cells using human umbilical vein endothelial cells by indirect co-culture. DESIGN, TIME AND SETTING: In vitro study of cell engineering was done at the Medical Research Center of Guangdong Provincial People’s Hospital between January and July 2007. MATERIALS: Small quantities of bone marrow were obtained from 11 children with congenital heart disease but without hematologic diseases through manubrium of sternum puncture in the congenital heart defect corrective surgery after the permission of family member. Umbilical cord of full-term normal delivery healthy newborn was provided by the Department of Obstetrics, the First Affiliated Hospital of Sun Yat-sen University. Fresh cattle jugular vein was provided by Guangdong Dali Meat Cattle Butchery. METHODS: The hBMSCs were isolated and purified using density gradient centrifugation method and were cultured in vitro. Human umbilical vein endothelial cells were obtained from newborn umbilical cord by enzyme digestion. Cell culture insert with semipermeable membrane combined with 6-well plate was used to do indirect co-culture induction. Human umbilical vein endothelial cells were expanded in the cell culture insert, passage 3 bone marrow mesenchymal stem cells were expanded in the 6-well plate outside the culture insert at the density of 1?105 cells/well, the initial ratio of the two kinds of cells was 1:5, then low-glucose DMEM culture solution containing 10% fetal bovine serum was added, cells were cultured for 14 days. Co-culture of bone marrow mesenchymal stem cells and bone marrow mesenchymal stem cells was used as control. Introduced endothelioid cells were cultured and then seeded on the cell-free cattle jugular vein intravascular stent. MAIN OUTCOME MEASURES: Morphology changes of induced cells; introduced endothelioid cell surface antigen detected through immunocytochemical staining; the growth and adhesion condition of endothelioid cells on the intravascular stent observed under scanning electronic microsope. RESULTS: The morphologies of introduced endothelial cells were uniform, introduced endothelial cells presented a cobblestone-like appearance, they amplified fast and expressed endothelial cell-specific surface marker CD31 and vWF and the positive rate was over 99%. They also could form a continuous unicellular layer on the cell-free cattle jugular vein intravascular stent. CONCLUSION: Human umbilical vein endothelial cells can induce hBMSCs to differentiate into endothelial cells successfully and to adhere and grew on the cell-free cattle jugular vein intravascular stent through indirect co-culture method.
