Results of selective isolation of NK cells from peripheral blood

Abstract

Background: Natural killer (NK) cells, which compose 5 to 20 percent of peripheral blood lymphocytes, are the first to recognize and destroy cancer cells and virus infected cells. The basic principle of NK cell immunotherapy is to isolate NK cells and activate them with anti-inflammatory cytokines to transfuse them into the recipient. Purity of isolated NK cells is directly related to the effectiveness of immune cell therapy. Immune cell therapy is practiced in many countries around the world, and the selective isolation of NK cells will be part of the basic research on the introduction of immune cell therapy in our country. There is no study on the selective isolation of immune cells in our country. Aim: To do selective isolation of NK cells from peripheral blood using microbeads Materials and methods: Peripheral blood from volunteers was used as a test sample. Mononuclear cells were isolated from peripheral blood using a Ficoll-Paque PLUS solution with a difference in density. NK cells were isolated from mononuclear cells using a CD56 magnetic bead (CD56 MicroBeads, human, Miltenyi Biotec, Germany) by MACS (Magnetic Activated Cell Sorting, MACS, Miltenyi Biotec, Germany). The isolated NK cells were placed on a slide glass using a cytospin centrifuge, fixed and stained with a MayGrunwald (Cypress Diagnostics, Belgium) and evaluated under a microscope. Cell counts were determined using Trypan blue dye (Trypan blue dye, 0.4% solution, Bio-Rad Laboratories, USA) and an automated cell counter (Bio-Rad TC20TM Automated Cell Counter, USA). Results and conclusion: Mononuclear cells with a density difference of 4.01x106 cells/ml (of which the number of viable cells was 2.57x106 cells/ml and the percentage of viable cells was 61%) were isolated from 3ml of peripheral blood using ficoll solution. Approximately 5.04x104 cells/ml of CD56 positive NK cells (of which 3.27x104 cells/ml, 53% of which were viable cells) were isolated from of 4.01x106 mononuclear cells. Approximately 5.04x104 cells/ml (viable cells 3.27x104 cells/ml, 53%) of CD56 positive NK cells can be selectively isolated from 3 ml of peripheral blood using CD56 microbeads.