Vacutainer pst

We generated lymphoblastoid cell lines (LCLs) from the affected member and both of his parents, by using the previously described method [68 (link)]. In short, blood samples collected in lithium heparin tubes (BD Vacutainer^® PST™) were lysed in buffer (155 mM ammonium chloride, 10 mM potassium hydrogen carbonate and 0.1 mM disodium-EDTA) for 10 min. Immortalisation of lymphocytes was performed by Epstein-Barr-Virus and to remove T-lymphocytes, cyclosporine was used. This resulted in the generation of only B-lymphocytes.

All sample collection was performed following signed and informed consent prior to invasive procedure and sample testing as outlined by the TCPS2. Samples were alphanumerically coded and sample testing was performed single blinded. A small sample of venous blood (2 mL) is collected from both athletes and non-athletes into lithium heparin coated vacutainer tubes (BD vacutainer PST, #367962). Samples are immediately placed on ice to allow stable transport to laboratory setting. Samples were then centrifuged at 10,000G for 10 minutes for plasma isolation. Plasma fraction was aliquoted and immediately stored at-80^°C for future analysis.

We generated lymphoblastoid cell lines (LCLs) from subjects 1 and 2 from blood samples collected in lithium heparin-coated tubes (BD Vacutainer PST). Red blood cells were lysed through incubation in lysis buffer (155 mM ammonium chloride, 10 mM potassium hydrogen carbonate, and 0.1 mM disodium-EDTA) for 10 min. Pelleted lymphocytes were immortalized by transfection of Epstein-Barr virus, followed by incubation at 37 °C for 1 h. Finally, cyclosporine was added to selectively kill T lymphocytes and generate only a cell line of B lymphocytes as detailed previously.² (link) In short, LCLs were grown in RPMI 1640 Medium, GlutaMAX Supplement (61870044; Thermo Fisher Scientific), supplemented, and fortified with 10% fetal bovine serum (FBS; Biochrom), L-glutamine (P04-80050; PAN Biotech), and antibiotics (penicillin/streptomycin, P06-07050; PAN Biotech). Cells were cultured at 37 °C in incubators supplied with 5% CO₂.