Gravity gradient sedimentation: A method for preparation of single cell suspension of varying densities from tissues

Single cell from various fetal and adult tissues of mouse, rat and human were isolated according to their densities by gravity gradient sedimentation method. In this method, tissues were minced in a suspension medium and gently forced through four nylon bloting clothes of decreasing pore sizes, without centrifugation. The cells were then allowed to distribute themselves in a exponential gradient solution according to their densities and no external centrifugal force (g) was applied. The gradient was then collected in two fractions and centrifuged. The intact cells of different densities were then utilized as an antigen fm:several immunological techniques including enzyme linked immunosorbent assay. This method was well suited for checking cross reactivity and tissue speciticity of mouse monoclonal antibodies to several tissues of different histological origin and for different stage of embryonic development by ELISA. Isolation of single cells by their mechanical dispersion method without destroying the cell membrane bound antigens were proved more suitable over the trypsinization method using ficoll gradient.