EBV Immortalization of human B lymphocytes separated from small volumes of cryo-preserved whole blood.
نویسندگان
چکیده
EBV immortalized B lymphocyte cell lines have been extensively used as a source of biological material for functional and molecular studies and represent a potentially limitless source of genomic DNA. Current technologies for EBV transformation are costly and use relatively large volumes of peripheral blood. Alternative methods were examined to determine whether smaller volumes of cryo-preserved whole blood could be subsequently transformed and which could provide a more cost-effective strategy for large population-based studies such as UK Biobank. A successful method was established where viable B cells were positively selected from 0.5 ml cryo-preserved whole blood samples. These were EBV transformed in microtitre plates and subsequently expanded in culture. A pilot study within UK Biobank was performed, which confirmed its potential usefulness for this study.
منابع مشابه
Generation of cell lines using Epstein-Barr Virus (EBV) transformation of small volumes of cryo-preserved whole blood and the use of bench-top flow cytometry to achieve high and reproducible success
ECACC has developed a procedure for the generation of lymphoblastoid cell lines (LCLs) by the direct EBV transformation of small amounts (800 l) of cryo-preserved whole blood eliminating any requirement for prior separation of peripheral blood lymphocytes (PBLs). The ability to directly transform frozen whole blood avoids the cost of separating and storing PBLs while retaining the option to mak...
متن کاملEffect of Myeloma-Derived Microparticles on in Vitro Proliferation and Viability of Alloimmune Human Peripheral Blood Mononuclear Cells
Background: Microparticles (MPs) are small vesicles released from the cell membrane. Accordingly, these contain active molecules to mediate biological processes, including cell proliferation and cell cycle progression. The fusion of myeloma cell lines with immunized B lymphocytes is a critical step of hybridoma technology. MPs modulate lymphoproliferation, thereby facilitating B cell expansion ...
متن کاملMycoplasma fermentans infection promotes immortalization of human peripheral blood mononuclear cells in culture.
Chronic infection or colonization by mycoplasma(s) could gradually and significantly alter many biologic properties of mammalian host cells in culture, including induction of malignant transformation. We examined effects of Mycoplasma fermentans infection on the continuing survival and immortality of human peripheral blood mononuclear cells (PBMCs) from healthy blood donors. Without specific su...
متن کاملPhysiologic concentrations of normal human plasma lipoproteins inhibit the immortalization of peripheral B lymphocytes by the Epstein-Barr virus.
Epstein-Barr virus (EBV)-induced immortalization of adult human B lymphocytes is suppressed by physiologic concentrations of human plasma lipoproteins. Several inhibitory mechanisms appear to be operative. First, low density lipoproteins (LDL) directly reduce the ability of EBV to transform human B cells. Second, LDL as well as intermediate and very low density lipoproteins modulate early induc...
متن کاملPersistent infection of Epstein-Barr virus-positive B lymphocytes by human herpesvirus 8.
In patients with Kaposi's sarcoma (KS), human herpesvirus 8 (HHV-8) can invariably be detected in KS tumor tissue and, at a lower frequency, in prostate tissue and peripheral blood B lymphocytes. Whereas the majority of KS spindle cells are latently infected by HHV-8, linear HHV-8 genomes characteristic for lytic infection are found predominantly in the peripheral blood cells of KS patients. In...
متن کاملذخیره در منابع من
با ذخیره ی این منبع در منابع من، دسترسی به آن را برای استفاده های بعدی آسان تر کنید
عنوان ژورنال:
- International journal of epidemiology
دوره 37 Suppl 1 شماره
صفحات -
تاریخ انتشار 2008