| 摘要 |
PURPOSE:To provide a gene induced from a mRNA induced in a myelocytic cell stimulated with a granulocyte colony-stimulating factor (G-CSF) and used for the detection of the myelocytic cell, for a sensitivity test to the G-CSF, etc. CONSTITUTION:A bone marrow is collected from the ileum of a chronic myelocytic leukemic patient of the chronic period with a heparin injector using a bone marrow- sticking needle under the lumbar anesthesia of the patient and subsequently added to a medium. Myelocytic cells such as monocytes are collected from an intermediate layer produced by a specific gravity centrifugal method, cultured in a medium containing 10% fetal calf serum and the granulocyte colony-stimulating factor (G-CSF) in an amount of 50ng/ml in a culture device maintained at 37 deg.C and a 5% CO2 content for 18 hr, and subsequently centrifuged to obtain the cell pellets. The cell pellets are suspended in a buffer solution, ground with a cell-grinding machine and then subjected to the separation of all RNAs by a conventional method. All RNAs are treated with an oligo dT-bonded carrier to separate mRNA, and the mRNA is treated with a reverse transcriptase to synthesize cDNA, thus obtaining the cDNA for the mRNA induced from the myelocytic cells stimulated with the G-CSF. |
