・         Lymphocyte Transformation (Donis-Keller lab) Lymphocytes are transformed to establish cell lines. Mononuclear cells (lymphocytes) from anticoagulated venous blood are isolated by layering onto histopaque. During centrifugation, erythrocytes and granulocytes are aggregated by ficoll and rapidly settle to the bottom of the tube; lymphocytes and other mononuclear cells remain at the plasma-histopaque interface. Erythrocyte contamination is neglible. Most extraneous platelets are removed by low speed centrifugation during the washing steps.

・         Preparation of Lymphoblastoid Cell Lines for Long-Term Storage (Donis-Keller lab) Cell are frozen in RPMI-1640 with 15% Fetal Bovine Serum + 10% DMSO. Cultures are frozen slowly using a Model 700 Controller freezing chamber... 

・         Maintaining Lymphoblastoid Cell Lines (Donis-Keller lab) To grow lymphoblastoid cells for permanent storage and for DNA extraction.

・          Lymphoblastoid Cell Lines from Frozen Whole Blood (Donis-Keller lab) Blood Samples can be stored frozen as a backup in case an LCL is needed at a later date.   

• Preparation of Lymphocyte Cell Pellet for Storage (Donis-Keller lab) Following propagation to 1 X 108 cells, lymphoblastoid cells are conveniently stored at -80 degrees C to preserve the high molecular weight DNA in the cells until the DNA is purified. This procedure describes the steps required to harvest and freeze the cells for long term storage.   
• CD14 CD16 Staining Protocol (Ziegler-Heitbrock)   
• Phagocytosis (Devreotes Lab)   
• Needle Assay for Chemotaxis (Devreotes Lab)