Preliminary Isolation Of Renal Carcinoma Stem Cells And Their Biological Characteristics

Objective 1.To explore the method of primary culture,purification and separation of renal carcinoma cells, and to establish the experimental patterns for studying renal carcinoma cells. 2.To separate renal carcinoma stem cells (RCSC) and study their general biological characteristics.Methods Fresh renal carcinoma tissue was made into single cell suspension by mechanical and enzymatic dissociation, and was purified with isopycnic sedimentation centrifugation and repeated adherence, then CD133 immunomagnetic beads were used to separate the single cell suspension, CD133~+ cells and CD133~-cells harvested were cultured separately to determine clone formation and reproductive activity; simultaneously, renal tumor CD133~+ cells and CD133~- cells were subcutaneously injected in SCID mice to assess their potential ability to form tumor clone; and describe their morphologic features.Results Conditioned medium for primary culture,purification and isolation of renal carcinoma cells was neccessary; Clone formation and reproductive activity of CD133~- cells are stronger than CD133~+ cells in our experiment, and all of the CD133~+cells,CD133~- cells and the cells before separation did not form tumor in our four cases of subcutaneous implantation in SCID mice.Conclusions Constitution of the renal carcinoma cells is complicated. Renal carcinoma stem cells are possibly existed in subpopulation of CD133 positive cells.