Introduction
How to use the Human Genetic Cell Bank Service
Do I pay to receive the cell lines once generated?
Can I have DNA instead of cell lines?
Processing of Blood Samples at the Human Genetic Cell Bank
Authentication of Cell Lines
Acknowledgment of Receipt of Blood Samples
Will I receive regular updates on my samples?
Guidelines Regarding the Collection and Postage of Samples
Blood Samples
Peripheral Blood Lymphocytes (PBLs)
General Postage & Packaging of Samples
What is the cost of the service?
Introduction
The Human Genetic Cell Bank Service at ECACC is based on Epstein-Barr Virus (EBV) immortalisation of human peripheral blood lymphocytes (PBLs) resulting in the production of lymphoblastoid cell lines (LCLs). These cell lines are subsequently banked, providing a renewable and expandable source of genomic DNA and RNA. This service is available to both academic and commercial organisations.
Although ECACC has continued to receive funding from the Medical Research Council (MRC) we are no longer funded to provide a subsidised service for processing blood samples to PBLs and/or establishing EBV transformed lymphoblastoid cell lines. The published price list therefore reflects the full operational costs of providing these services and is applicable to all users of the service. There is no prerequisite to make subsequent cell lines available through the published ECACC catalogue although we would actively encourage this in order to improve genetic resources available for research. However, should there be any concerns in doing this then access to your deposits can be restricted to reflect your requirements.
How to use the Human Genetic Cell Bank Service
Depending on your requirements there are two routes to using the Human Genetic Cell Bank Service. You may be about to embark on a large study for which you intend to deposit multiple samples (generally in excess of 50 samples), alternatively you may wish to send just one or a few individual samples (less than 50 samples during a year). Please refer to (a) or (b) below according to your needs:
(a) For Studies (usually in excess of 50 samples):
Contact ECACC directly to discuss your requirements or submit a completed Genetic Study Registration Form to ECACC. This form requests details of the disorder for which samples are to be collected along with the estimated number of samples involved, and your requirements. This will be used to compile a specific quotation.
Evidence of Research Ethics Committee approval and a sample copy of the detailed patient consent form must also be provided. A meeting with ECACC staff is encouraged prior to a project starting.
On written customer acceptance of the quotation a specific Statement of Work will be issued by ECACC including standard terms and conditions for using the service. A signed copy of this must be in place before ECACC can begin receiving samples.
Following acceptance of the study by ECACC each sample sent to ECACC must be accompanied by a completed Human Genetic Cell Bank Form. Alternatively, an equivalent form provided by the Principal Investigator of the study which has been approved by ECACC may be used.
(b) For Small Numbers of Samples (generally less than 50 samples during a year):
Contact ECACC directly to inform us of your intention to send a sample. Please do this prior to collecting samples so that we can prepare for the arrival of samples and provide advice on the best way to handle and send samples. Each sample must be accompanied by a completed Human Genetic Cell Bank (HGCB) Deposit Form. Please note that evidence of patient consent must be provided, refer to the HGCB Deposit Form where space is provided regarding patient consent.
Do I pay to receive the cell lines once generated?
Each depositor is entitled to obtain frozen vials of their deposits free of charge (excluding shipping). For a growing culture there is a start-up fee unless the cell line is requested at the time of transformation, in which case we can send you a flask of cells free of charge (excluding shipping).
In all cases, shipping charges for return of cell lines will be charged to the depositor at the current rate. Customers requesting cell lines deposited by someone else will be required to pay shipping charges.
Can I have DNA instead of cell lines?
Cell pellets for DNA extraction, purified DNA or RNA can be produced. Please enquire for a specific quotation.
Processing of Blood Samples at the Human Genetic Cell Bank
Peripheral blood lymphocytes (PBLs) are separated from the blood samples received. The isolated PBLs are frozen in two aliquots provided sufficient numbers are obtained and the blood (ideally 6-8 ml) is received in good condition soon after collection. Alternatively PBLs are transformed immediately if specifically requested by the depositor, or if:
(i) a low volume of blood, i.e. 4ml or less, is received
(ii) insufficient numbers of PBLs are extracted
(iii) the blood sample is of poor quality
In some instances, for example if PBL separation is not possible, the sample will be discarded and the depositor advised immediately by telephone or email.
Frozen PBLs are resuscitated from liquid nitrogen storage when scheduled to undergo EBV transformation. Test batches of transformations are performed for projects where only PBL storage is requested.
Lymphoblastoid cell lines (LCLs) generated are stored in the ECACC cryostore facility. Cell lines can be supplied as frozen ampoules or growing cultures. In addition, ECACC can extract and supply DNA and RNA from LCLs.
Figure: Overview of the Human Genetic Cell Bank EBV Transformation Service Authentication of Cell Lines
Bloodspot cards are kept for all blood samples received. These act as a reference for ECACC to verify that cell lines generated have the same identity as the original deposited blood sample.
Acknowledgment of Receipt of Blood Samples
For studies, reports listing all samples received are issued monthly, otherwise receipt of blood samples is acknowledged by post (usually within one week) informing the depositor of whether the sample has been transformed or stored as PBLs.
Depositors are invoiced monthly if they are regularly depositing samples. A purchase order number will be required from the depositor for this purpose.
Will I receive regular updates on my samples?
Monthly status reports are issued for all studies for which samples are being transformed. Status reports for individual samples will not normally be provided on a regular basis. However, if at any time you wish to have an update on any aspect of your deposit, contact ECACC directly. You will automatically be informed if samples fail to transform with an explanation, if known.
It should be appreciated that a number of factors, including age and disease condition of the donor can influence transformation success. In addition, approximately 2% of the population have lymphocytes that are not susceptible to transformation. Undoubtedly, the higher the quality of the blood sample the greater the potential for producing a cell line. Please refer to the information regarding the collection and postage of blood samples.
Guidelines Regarding the Collection and Postage of Samples
If you are planning to send either blood samples or PBLs you should contact ECACC to discuss your requirements and to arrange for a quotation. Please read the information on 'How to use the Human Genetic Cell Bank Service'. For larger projects we require a copy of the Research Ethics Committee approval for the project and a sample copy of the detailed patient consent form. If you only plan to send a small number of samples, contact ECACC prior to collection of samples and ensure a Human Genetic Cell Bank Deposit Form, including confirmation of informed donor consent, accompanies each sample.
Blood Samples
- Use acid citrate dextrose (ACD) or citrate phosphate dextrose adenine (CPDA) blood tubes and ensure proper mixing by gentle inversion of the tubes.
- All samples must be clearly labelled, preferably with a bar-code, and accompanied by a deposit form.
- Ideally 6-8ml of blood is required.
- Samples should be sent on day of collection, and should arrive at ECACC preferably the next day but certainly within 72 hours. This is particularly important for samples in Heparin or EDTA, as sample deterioration is marked after 48 hours in comparison to samples in ACD or CPDA tubes.
- Store samples at room temperature (15-21oC). DO NOT refrigerate or freeze under any circumstances.
- If a third party is sending blood on your behalf, please ensure that they are made aware of these recommendations.
- Please give prior warning of any samples which will arrive at ECACC at the weekend, so that provision can be made to process the samples.
- Samples must be packaged according to International Air Transport Association (IATA) safety regulations (full details on website, www.iata.org).
- Samples with a known HepB, HepC or HIV risk should not be sent without prior agreement. ECACC requires at least seven working days notice, as special arrangements have to be made to process such samples. In the absence of such notice, recognisably biohazardous samples will be destroyed on receipt.
Peripheral Blood Lymphocytes (PBLs)
- PBLs should be prepared in a dedicated, mycoplasma free area that is not used for routine cell culture or the processing of human pathology specimens. If the former is not possible, screening of cell cultures for mycoplasma contamination should be carried out on a regular basis. ECACC must be informed of any potential risk of mycoplasma contamination.
- PBLs should be frozen to <-80oC at a controlled rate, in the presence of cryoprotectant. Internal threaded cryovials, clearly labelled (preferably barcoded) with the sample reference on wrap-around, low temperature resistant labels, should be used. Do not use hand-written labels.
- Frozen PBLs should be sent to ECACC in an insulated carton, packed in dry ice sufficient for 3-5 days depending on the distance to travel.
- A blood spot card should be provided for each sample where possible.
- PBL shipments should be accompanied by a list of the samples and their identities, grouped to reflect the packaging. The identifiers should be identical to the cryovial labels. Affix reproductions of the barcodes if appropriate.
- At least 24 hours notice should be communicated to ECACC prior to shipment of frozen PBLs.
General Postage & Packaging of Samples
Since 1st April 1999 there have been new regulations regarding the shipment of infectious and biological samples. Failure to comply with these regulations may result in the prosecution of the sender. The regulations require that all infectious substances sent with the Royal Mail must comply with UN 602 packaging specifications. These are as follows.
The packaging system should consist of three layers:
o Primary receptacle
A labelled leak-proof container for the sample.
o Secondary receptacle
A durable, watertight receptacle to enclose and protect the primary container. Several primary containers may be placed in one secondary receptacle. Absorbent material, sufficient to retain the total volume of liquid, must be used to cushion the primary containers
o Outer shipping package
The secondary receptacle is placed in an outer shipping package which protects it from physical damage and water whilst in transit. Use additional absorbent material to cushion the secondary container
Approved UN 602 packaging may be obtained from several suppliers; details are available from any Royal Mail Sales Centre.
If you use a courier service to transport your blood samples, please contact the company involved for advice on their own approved packaging. ECACC will be happy to return any packaging to you for re-use.
What is the cost of the service?
The service is priced as follows:
Sample booking in (blood or frozen PBL) (*Stage 1) £5 per sample
Blood processing to frozen PBL (*Stage 1) £30 per sample
EBV immortalisation (*Stage 2) £80 per sample
If your project is funded by the Wellcome Trust you may eligible for a reduced price. Please enquire for a specific quotation.
*Refer to the figure which provides an overview of the Human Genetic Cell Bank EBV Transformation Service
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