ABSTRACT
Objective To investigate the immunologic properties of osteogenic differentiated bone mesenchymal stem cells (BMSCs). Methods BMSCs were isolated from normal volunteers and induced in osteogenic medium for two weeks. Then,non-differentiated/osteogenic differentiated BMSCs were co-cultured with allogenic T cells and phytohemagglutinin (PHA).The proliferation of T cells was examined by MTT method.The concentrations of TGF-β1 in osteogenic differentiated BMSCs supernatants at week 2 and mixed lymphocytes reaction (MLR) supernatants at day 5 were determined by ELISA.Also,anti-TGF-β antibody was added into the MLR to detect the response of the mixed T cells. Results Non-differentiated and osteogenic differentiated BMSCs did not induce proliferation of the allogeneic T cells but both suppressed the proliferation of the T cells mediated by PHA.The TGF-β1 concentrations had significant elevation in the MLR.Anti-TGF-β antibody could counteract the immunosuppressive function of the osteogenic differentiated BMSCs. Conclusion Osteogenic differentiated BMSCs possess low immunogenicity and immunosuppressive property.
ABSTRACT
BACKGROUND: Tissue engineering development brings a hope for articular cartilage defect repair. Current studies have demonstrated that bone marrow mesenchymal stem cells (MSCs) are the best cell source to repair full-thickness cartilage defects.OBJECTIVE: To induce MSCs to differentiate into chondrocytes with Ad-hTGF-pi in pellet culture system in vitro and identify the differentiated cells.DESIGN, TIME AND SETTING: Repetitive cellular measurements were performed at the Central Laboratory of Xinqiao Hospital,Third Military Medical University of Chinese PLA from June 2007 to January 2008.MATERIALS: Japanese rabbits, 2 months old, were provided by the Laboratory Animal Center of Third Military Medical University of Chinese PLA.METHODS: The rabbit MSCs were isolated, cultured and expanded in vitro. After transacted with Ad-hTGF-β1, the cells were cultured in pellet culture system. Chondrogenic differentiation was evaluated by histological, immunohistochemical and RT-PCR techniques.MAIN OUTCOME MEASURES: Cell morphological changes were observed by histological staining; proteoglycan and type Ⅱ collagen expression was detected by immunohistochemical and RT-PCR techniques.RESULTS: The induced cells exhibited a chondrocyte-like morphology by histological staining. Immunohistochemical and RT-PCR results showed that proteoglycan and type Ⅱ collagen were expressed in the induced cells.CONCLUSION: Bone marrow MSCs cultured in pellet culture system can differentiate into chondrocytes under the induction of Ad-hTGF-β1.
ABSTRACT
BACKGROUND:Tissue engineering development brings a hope for articular cartilage defect repair.Current studies have demonstrated that bone marrow mesenchymal stem cells (MSCs) are the best cell source to repair full-thickness cartilage defects.OBJECTIVE:To induce MSCs to differentiate into chondrocytes with Ad-hTGF-?1 in pellet culture system in vitro and identify the differentiated cells.DESIGN,TIME AND SETTING:Repetitive cellular measurements were performed at the Central Laboratory of Xinqiao Hospital,Third Military Medical University of Chinese PLA from June 2007 to January 2008.MATERIALS:Japanese rabbits,2 months old,were provided by the Laboratory Animal Center of Third Military Medical University of Chinese PLA.METHODS:The rabbit MSCs were isolated,cultured and expanded in vitro.After transfected with Ad-hTGF-?1,the cells were cultured in pellet culture system.Chondrogenic differentiation was evaluated by histological,immunohistochemical and RT-PCR techniques.MAIN OUTCOME MEASURES:Cell morphological changes were observed by histological staining;proteoglycan and type II collagen expression was detected by immunohistochemical and RT-PCR techniques.RESULTS:The induced cells exhibited a chondrocyte-like morphology by histological staining.Immunohistochemical and RT-PCR results showed that proteoglycan and type II collagen were expressed in the induced cells.CONCLUSION:Bone marrow MSCs cultured in pellet culture system can differentiate into chondrocytes under the induction of Ad-hTGF-?1.