In vitro reconstruction of tissue function with the immortalized cell lines from transgenic mouse

为了利用永生化组织细胞系在体外重建组织功能，我们从SV 40大T抗原基因（ts T）转基因小鼠中建立了具有分化功能的永生化细胞系，并将其与骨髓基质细胞共培养，建立了4株新的未成熟造血干细胞样细胞系。THS 119细胞具有与骨髓造血干细胞相似的Lin-，c-Kit+ Sca-1+表面表型。B31-1细胞系显示B淋巴祖细胞的特性。DFC细胞由Dexter型骨髓培养物产生，并且DFC-α（克隆分离物）显示类似于双能骨髓祖细胞的表型，并且DFC-28显示非常早期的淋巴祖细胞。所有的细胞系都需要基质细胞来维持，利用这些共培养系统，此外，还从ts T转基因小鼠中建立了新的血脑屏障内皮细胞系、肾上腺细胞系和脑星形胶质细胞系。建立了具有特殊组织功能的免疫细胞系，如肝细胞系、肾小管细胞系和来自多种组织-血液屏障的新型内皮细胞系。

To reconstruct the tissue functions in vitro with the immortal tissue cell lines, we established several immortalized cell lines with differentiated functions from the temperature-sensitive SV40 large T-antigen gene (ts T) transgenic mice.By coculturing the hematopoietic stem cells with the bone marrow stromal cell lines established form the ts T transgenic mice, 4 novel immature hematopoietic stem cell-like cell lines including were established. THS119 cell line exhibited Lin-, c-Kit+ Sca-1+ surface phenotype similar to bone marrow hematopoietic stem cells. B31-1 cell line show properties of B-lymphoid progenitors. DFC cells were generated from Dexter type bone marrow culture and DFC-a, a clonal isolate, showed the phenotype similar to bipotent myeloid progenitor cells and DFC-28 showed very early lymphoid progenitor cells. All cell lines required stromal cells for their maintenance.Using these coculture systems, genes for 2 new membrane proteins (SMAP-1 and CD47/IAP), and a new cytoskeletal protein (mys-PDZ) were cloned and their function was examined.In addition, novel endothelial cell lines from blood-brain barrier, adrenal gland cell lines, and brain astrocyte cell lines were established from the ts T-transgenic mice.Recently, ts T-transgenic rats were generated and the immortlised cell lines with specialized tissue functions such as hepatocytes, kidney tubule cell lines, and novel endothelial cell lines from several tissue-blood barriers were established.