Isolation of B-1 subtype B lymphocytes from human cord blood

B1 lymphocytes by having a large supply of CD5 as a marker have the capability of cell differentiation and distinguishing them into cells similar to macrophages B1 Cell. The aim of the study is to determine the possibility of separation and purification of this unique category from other cellular organizations of cord blood lymphocytes. 14 samples of the umbilical cord blood from normal delivery or elective cesarean were used. Dilutions of cells to purified circulating Leukocytes by the use of hydroxyl ethyl starch or 3% gelatin have been made and WBC was separated from RBC. In order to separate adhesive or non-adhesve cells، cell culture was done. By gradient ficoll and centrifuge، lymphocyte cells of the umbilical cord from the remaining leukocytes and mononuclear cells were purified and they were extracted from B lymphocytes by nylon wool columns. And rosette test was used for testing the extracted cells. In spite of lack of difference between cellular counts in the two blood dilution materials، the amount of cellular survival showed to be lower than 87% in hydroxyl ethyl starch method while in the dilution method with gelatin 3%، the cellular life was about 90-95%. The results of Rosette test showed that dilution amount of B1 cells by the above method was about 99%. This process reveals that most of B cells extracted from cord blood are the of B1 cell type and the best and most suitable technique is gelatin 3% procedure with regard to the number and amount of cellular’s life.