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Expression of IgH promoter/enhancer Ly-1 transgene in hematopoietic chimeric mice generated by embryonic stem cell
1Second Department of Internal Medicine, School of Medicine, Chiba University, Japan.
We have produced hematopoietic chimeric mice from an embryonic stem (ES) cell line carring Ly-1 cDNA under the control of IgH promoter and enhancer. Various amounts of serum IgM (5-86% of total IgM) in chimeric mice were of ES origin and 30-60% of IgM-positive B cells from the chimeric mice analyzed were reconstituted from ES cells. Using these chimeric mice, the expression of the Ly-1 transgene on lymphoid tissues was examined by polymerase chain reaction assay with primers specific for the transgene, and by cell sorter analysis. Transcription of the Ly-1 transgene was detected in spleen cells, thymocytes and lymph node cells; however, the expression of the Ly-1 molecule was observed only on lipopolysaccharide (LPS)-stimulated splenic IgM-positive B cells but not on resting splenic B cells. There was no significantly increased expression of Ly-1 on splenic T cells and thymocytes. Thus, our findings demonstrate that conventional splenic B cells could express the Ly-1 transgene on their surface in vivo after LPS stimulation. Also discussed is the ES-derived chimeric hematopoietic system.
We have produced hematopoietic chimeric mice from an embryonic stem (ES) cell line carring Ly-1 cDNA under the control of IgH promoter and enhancer. Various amounts of serum IgM (5-86% of total IgM) in chimeric mice were of ES origin and 30-60% of IgM-positive B cells from the chimeric mice analyzed were reconstituted from ES cells. Using these chimeric mice, the expression of the Ly-1 transgene on lymphoid tissues was examined by polymerase chain reaction assay with primers specific for the transgene, and by cell sorter analysis. Transcription of the Ly-1 transgene was detected in spleen cells, thymocytes and lymph node cells; however, the expression of the Ly-1 molecule was observed only on lipopolysaccharide (LPS)-stimulated splenic IgM-positive B cells but not on resting splenic B cells. There was no significantly increased expression of Ly-1 on splenic T cells and thymocytes. Thus, our findings demonstrate that conventional splenic B cells could express the Ly-1 transgene on their surface in vivo after LPS stimulation. Also discussed is the ES-derived chimeric hematopoietic system.