Ficoll hypaque density gradient

A total of 42 plasma samples were collected from DLBCL patients diagnosed from 2017 to 2019 at Shandong Provincial Hospital. Plasma from 31 volunteers were randomly collected at the same period as controls. Informed consent was obtained from each patient. Peripheral blood mononuclear cells (PBMCs) were isolated by the Ficoll-Hypaque density gradient centrifugation method (TBD Science, Tianjin, China). The study was abided by Helsinki protocol and approved by the Medical Ethical Committee of Shandong Provincial Hospital.
DLBCL cell lines OCI-LY1, OCI-LY3, OCI-LY8, and OCI-LY10 were purchased from American Type Culture Collection (ATCC, Manassas, VA, United States). Cells were cultured in IMDM with 10% fetal bovine serum (FBS, GIBCO, MD, United States) and 1% double antibiotics, incubated at 37°C in 5% CO₂. All cells were examined for mycoplasma infection periodically.

This study was approved by the Shandong Provincial Hospital Affiliated to Shandong University Medical Ethics Committee with the informed consent of each patient based on the Helsinki Declaration. 51 CLL patients newly diagnosed according to the International Workshop on Chronic Lymphocytic revised criterion were collected from January 2013 to September 2019. All samples were obtained following informed consent. The peripheral blood mononuclear cells of patients were extracted by Ficoll-Hypaque density gradient (TBD Science, Tianjin, China). And normal CD19 + B cells were extracted by CD19 + magnetic microbeads kit (Miltenyi Biotec, Bergisch Gladbach, Germany) according to the method previously reported [12 (link), 28 (link)].