中国病理生理杂志2011，Vol.27Issue(6)：1193-1198,6.DOI:10.3969/j.issn.1000-4718.2011.06.027
人AGM区及胎肝基质细胞促进小鼠胚胎干细胞向造血干细胞分化和体内造血重建
Supportive effects of human AGM region and fetal liver stromal cells on differentiation of murine embryonic stem cells into hematopoietic stem cells and in vivo hematopoietic reconstitution
摘要
Abstract
AIM: To investigate the differentiation of murine embryonic stem cells ( ESCs ) into hematopoietic stem cells ( HSCs ) by the supportive effects of human aorta - gonad - mesonephros ( AGM ) region and fetal liver ( FL ) stromal cells.METHODS: E14 ESCs were induced into embryoid body ( EB ) first.Then the cells from EB were further co - cultured with human AGM region and FL stromal cells in non - contact system.On day 6, the cells derived from EB were collected for Sea - 1 + c - Kit+ cell analysis by flow cytometry, colony forming unit ( CFU ) assay and teratoma formation checking.BALB/c female mice conditioned with lethal dose of [60Co]γ - ray irradiation were transplanted with EB cells from different culture systems.The survival rates, engraftment of donor cells, reconstitution of hematopoietic were monitored.RESULTS: Sea - 1 + c - Kit+ cells in EB cells co - cultured with human AGM region and FL stromal cells had the value of ( 21.96 ± 2.54 )％ , and the total CFU was as ( 520 ± 52 )/105 cells, which were statistically greater than those in EB cells only cultured with human AGM region stromal cells ( P ＜0.05 ).No teratoma was found in NOD - SCID mice after subcutaneous injection of EB cells co - cultured with human AGM region and FL stromal cells.In BALB/c female mice transplanted of EB cells co - cultured with human AGM region and FL stromal cells, the survival rate was 77.8％ ,and the peripheral blood cell count was obviously improved on day 14.PCR results showed the recipients all had sry gene copies from donor in bone marrow.The recipient mice transplanted with EB cells only cultured with human AGM region stromal cells all died within 15 days.CONCLUSION: Stromal cells from human AGM region and FL enhance the directed differentiation of ESCs into HSCs which can reconstruct hematopoiesis in vivo.关键词
主动脉-性腺-中肾区/肝/基质细胞/胚胎干细胞/造血复建Key words
Aorta-gonad -mesonephros region/ Liver/ Stromal cells/ Embryonic stem cells/ Hematopoietic reconstitution分类
医药卫生引用本文复制引用
蔡耘,张绪超,陈惠芹,吴北燕,黄绍良..人AGM区及胎肝基质细胞促进小鼠胚胎干细胞向造血干细胞分化和体内造血重建[J].中国病理生理杂志,2011,27(6):1193-1198,6.基金项目
国家自然科学基金资助项目(No.30300377) （No.30300377）
