Tissue Culture (B cells)

Location: Eastlake Building, E1-305
Contact phone: (206) 667-3756
Contact fax: (206) 667-2892
Contact e-mail: rcb@fredhutch.org

Tissue Culture is performed on human B-cells for the production and/or maintenance of B-lymphoblastoid cells (B-LCL). Peripheral Blood Mononuclear Cells (PBMC) can be provided or can be isolated from whole blood. Samples are then transformed into B-LCL using Epstein Barr Virus (EBV) to immortalize the cells and Cyclosporine A (CSA) to inhibit the T cell population. The B cells will undergo virally-induced transformation within 3 weeks of infection and the culture will be maintained until the desired quantity of cells has been reached for cryopreservation. Cells are frozen at 5-7 million cells/nunc using 10% DMSO in culture media. B-LCL can also be cultured for DNA isolation and extraction.

All cultures are done without the use of antibiotics.  The use of antibiotics can mask issues, leading to an incorrect determination of sample sterility and success.  During cryopreservation, a sample is obtained for downstream mycoplasma and bacteria contamination testing. Fungal testing is also available upon request.

Research Cell Bank also cultures B cells for maintenance of the Transplant Genomics Repository and the HLA reference standards.

Doing Business with Specimen Processing/Research Cell Bank

Eastlake Building, E1-305
SPL: (206) 667-4645
RCB: (206) 667-3756

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