AIM: To investigate the differentiation of rat bone marrow stem cells in liver after partial hepatectomy. METHODS: Bone marrow cells were collected from the tibia of rat with partial hepatectomy, the medial and left...AIM: To investigate the differentiation of rat bone marrow stem cells in liver after partial hepatectomy. METHODS: Bone marrow cells were collected from the tibia of rat with partial hepatectomy, the medial and left hepatic lobes were excised. The bone marrow stem cells (Thy^+CD3^-CD45RA^- cells) were enriched from the bone marrow cells by depleting red cells and fluorescence-activated cell sorting. The sorted bone marrow stem cells were labeled by PKH26-GL in vitro and autotransplanted by portal vein injection. After 2 wk, the transplanted bone marrow stem cells in liver were examined by the immunohistochemistry of albumin (hepatocyte-specific marker). RESULTS: The bone marrow stem cells (Thy^+CD3^-CD45RA^- cells) accounted for 2.8% of bone marrow cells without red cells. The labeling rate of 10 μM PKH26- GL on sorted bone marrow stem cells was about 95%. There were sporadic PKH26-GL-labeled cells among hepatocytes in liver tissue section, and some of the cells expressed albumin. CONCLUSION: Rat bone marrow stem cells can differentiate into hepatocytes in regenerative environment and may participate in liver regeneration after partial hepatectomy.展开更多
Previous studies have suggested that the transforming growth factor-β receptor ALK5 is crucial for articular chondrogenesis by bone marrow mesenchymal stem cells. Here, the wild-type ALK5 plasmids were mutated by ove...Previous studies have suggested that the transforming growth factor-β receptor ALK5 is crucial for articular chondrogenesis by bone marrow mesenchymal stem cells. Here, the wild-type ALK5 plasmids were mutated by overlapping extended PCR and transfected into bone marrow mesenchymal stem cells. The knee joint osteoarthritis mouse model was constructed by cutting oft" the anterior cruciate ligament and divided into three groups: saline group, bone marrow mesenchymal stem cells and ALK5-transfected bone marrow mesenchymal stem cells group. HE staining showed that the articular cartilage lesions were more serious of saline group compared with that of mesenchymal stem cell group, and this trend was more pronounced as time goes on. Immunohistochemical staining showed that although the expression level of type II collagen in all three groups down-regulated gradually upon time, its expression in ALK5-transfected bone marrow mesenchymal stem cells group was significantly enhanced compared with the other two groups. Micro-CT also suggested that ALK5 transfection of mouse bone marrow mesenchymal stem cells would promote repairing the knee cartilage lesions with arthritis of the mice. Although the osteoarthritis mechanism underlying a variety of factors work together, and the appropriate proportion of ALKS/ALK1 was also emphasized for the treatment of osteoarthritis. This work therefore demonstrated that ALK5 transfection of bone marrow mesenchymal stem cells could be a promising stem cell therapy for repair of cartilage lesions.展开更多
基金Supported by the National High Technology Research And Development Program of China (863 Program), No. 2001AA217151
文摘AIM: To investigate the differentiation of rat bone marrow stem cells in liver after partial hepatectomy. METHODS: Bone marrow cells were collected from the tibia of rat with partial hepatectomy, the medial and left hepatic lobes were excised. The bone marrow stem cells (Thy^+CD3^-CD45RA^- cells) were enriched from the bone marrow cells by depleting red cells and fluorescence-activated cell sorting. The sorted bone marrow stem cells were labeled by PKH26-GL in vitro and autotransplanted by portal vein injection. After 2 wk, the transplanted bone marrow stem cells in liver were examined by the immunohistochemistry of albumin (hepatocyte-specific marker). RESULTS: The bone marrow stem cells (Thy^+CD3^-CD45RA^- cells) accounted for 2.8% of bone marrow cells without red cells. The labeling rate of 10 μM PKH26- GL on sorted bone marrow stem cells was about 95%. There were sporadic PKH26-GL-labeled cells among hepatocytes in liver tissue section, and some of the cells expressed albumin. CONCLUSION: Rat bone marrow stem cells can differentiate into hepatocytes in regenerative environment and may participate in liver regeneration after partial hepatectomy.
基金This work was financially supported by National Natural Science Foundation of China (81600838, 51502262), Key Research and Development Program of Zhejiang, China (2017C01054), Medical Technology and Education of Zhejiang Province of China (2016KYB178), Research Science and Technology Department of Zhejiang Province social welfare development projects under Grant 2013C33161, Natural Science Foundation of Zhejiang Provence of China under Grant LY 15H 160019, and Ningbo Natural Science Foundation 2016A610166.
文摘Previous studies have suggested that the transforming growth factor-β receptor ALK5 is crucial for articular chondrogenesis by bone marrow mesenchymal stem cells. Here, the wild-type ALK5 plasmids were mutated by overlapping extended PCR and transfected into bone marrow mesenchymal stem cells. The knee joint osteoarthritis mouse model was constructed by cutting oft" the anterior cruciate ligament and divided into three groups: saline group, bone marrow mesenchymal stem cells and ALK5-transfected bone marrow mesenchymal stem cells group. HE staining showed that the articular cartilage lesions were more serious of saline group compared with that of mesenchymal stem cell group, and this trend was more pronounced as time goes on. Immunohistochemical staining showed that although the expression level of type II collagen in all three groups down-regulated gradually upon time, its expression in ALK5-transfected bone marrow mesenchymal stem cells group was significantly enhanced compared with the other two groups. Micro-CT also suggested that ALK5 transfection of mouse bone marrow mesenchymal stem cells would promote repairing the knee cartilage lesions with arthritis of the mice. Although the osteoarthritis mechanism underlying a variety of factors work together, and the appropriate proportion of ALKS/ALK1 was also emphasized for the treatment of osteoarthritis. This work therefore demonstrated that ALK5 transfection of bone marrow mesenchymal stem cells could be a promising stem cell therapy for repair of cartilage lesions.