Human Genetic Support Services
16 October 2007
Following extensive development, the ECACC can now offer its customers a service for the cryopreservation and EBV transformation from whole blood. Although transformation from separated lymphocytes remains the 'gold standard' method, cryopreservation of aliquots of whole blood for possible future EBV transformation is a viable alternative you may wish to consider.
Genomic DNA can be extracted from whole blood, fresh or frozen, using the latest automated technologies. The service is offered for blood volumes up to 10ml and includes quality control of DNA by agarose gel electrophoresis, quantification of DNA using the PicoGreen assay, and normalisation to a standard concentration. Final DNA can be supplied in a variety of formats, either plates or tubes, according to your specifications.
PERIPHERAL BLOOD LYMPHOCYTE (PBL) PREPARATION AND STORAGE
PBLs are separated from blood samples using density gradient centrifugation and cryopreserved in ECACC's ultra-low temperature storage facility. Blood spot cards for verification of cell line and DNA authenticity are prepared from every sample received. Cryopreservation of PBLs does not affect their subsequent EBV transformation; however, under certain conditions PBLs can be transformed immediately, without prior freezing.
Processing capacity is in excess of 1,500 blood samples per month, and samples are processed six days per week. ECACC is licensed under the Human Tissue Act 2004 for the storage of primary human material for research purposes.
LYMPHOBLASTOID CELL LINES
Lymphoblastoid cell lines (LCLs) are generated by Epstein Barr Virus (EBV) transformation of the B-lymphocyte component within the PBL population. Once a cell line has been successfully established, a bank of four ampoules is generated and stored in the ECACC Cryostorage facility. The process takes four to six weeks to complete. As part of the service, frozen cells or a growing culture can be returned to the customer.
We operate a 'right first time' policy and aim to achieve a transformation success rate of at least 95% at first attempt. We have the capacity to initiate 1,000 scheduled EBV transformations per month, the overwhelming majority of which are from frozen PBLs resuscitated from liquid nitrogen storage.
CELL CULTURE EXPANSION FOR DNA AND RNA EXTRACTION
We are able to generate frozen cell pellets to customer specifications from any of the cell lines available in our catalogue, in addition to your own deposit. Cell lines can be expanded to any volume of culture specified by the customer, however, for convenience, the majority of our customers choose frozen cell pellets prepared from three standard culture volumes.
Cell cultures can be utilised for DNA and/or RNA extraction and our protocols have been specifically designed to suit the differing requirements, in terms of point of cell harvest, reagents used, and freeze methods.
ECACC also offers a comprehensive contract cell culture service, combining a wide range of cell culture systems with extensive cell culture expertise, to address more specialised customer requirements.
EXTRACTION OF GENOMIC DNA FROM CELL LINES
High quality genomic DNA can be prepared from any cell line in ECACC's catalogue or your own collection, in quantities ranging from a few micrograms extracted from a frozen ampoule of cell culture, to milligram quantities, depending upon your needs. All DNA is quantified using the PicoGreen assay, and normalised to a standard concentration of 100ngìl-1 and quality verified by agarose gel electrophoresis. The final DNA can be supplied in a variety of formats, both plates and tubes, according to your specifications.
More information regarding the ECACC and its services can be found via the link below.
