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ECACC Handbook

Cell Culture

ECACC Handbook – Fundamental Techniques and Protocols for ECACC Cell Lines



1.0 The ECACC and its Collections

ECACC and Human Genetic Research

The European Collection of Cell Cultures (ECACC) was first established within CAMR, Salisbury, UK in 1984 and has grown to be one of the largest animal cell culture collections in the world. ECACC now supplies authenticated cell lines worldwide and provides a wide range of cell culture services.

Whereas the ECACC core business is cryopreservation, cell banking and the supply of authenticated cell lines the operation has progressively diversified and now offers a comprehensive range of services to cell scientists both in the commercial and academic sectors. These services include Patent and Safe deposits, screening for microbiological contaminants, authentication by DNA profiling and a comprehensive program of training courses. ECACC also undertakes contract cell culture process upscale development and the manufacture of cell products for research use.

The Human Genetic Collection

The fastest-growing service within the ECACC range is its Human Genetic Collection. This is based on processes introduced into, and subsequently developed by ECACC in the late 1980’s, for the immortalization of human lymphocytes. This process essentially comprises the separation of lymphocytes from samples of human peripheral blood and their transformation into lymphoblastoid cell lines by treatment with the tumourigenic Epstein Barr Virus (EBV). EBV transformation of human lymphocytes to form a cell line that can be continuously propagated provides an indispensable means of expanding the limited amount of cellular material that can be separated from a small volume blood specimen. This expanded cell stock can be banked and cryopreserved as a source of single donor cells and / or extracted DNA that can be made available indefinitely.

Since 1989 a number of major human genetic research programs, such as the British Diabetes Association-funded Warren I and II studies and the long-running, MRC-funded Human Genome Mapping Project, have depended upon the ECACC EBV transformation service. A cell line manufactured from a phenotypically characterized donor, who possibly has a rare genetic disorder, can be used to provide an almost limitless supply of DNA for distribution to research groups worldwide. Furthermore a cryopreserved bank of these cells can be resuscitated many years later so that investigation may be resumed with the benefit of improved technologies and scientific knowledge.

The ability to expand the amount of cellular DNA available from a small donor sample means that scientific investigations will not be materials limited. It is theoretically possible to achieve this expansion by biochemical means such as "whole genome PCR", but to date this has not proved practicable. Cell line DNA is typically of high quality and a viable cell offers the opportunity to examine other types of molecule, such as RNA or cellular proteins, if required at a later date.

The ECACC Human Genetic Collection now comprises more than 40,000 single donor lymphocyte stabilates, more than 50% of which are transformed cell lines. This collection represents more than 700 clinical disorders such as breast cancer, Alzheimers disease, schizophrenia and psoriasis.

The ECACC General Collection

This collection comprises approximately 1000 different cell lines originating from over 45 different species and a variety of different tissues. It incorporates over 400 human cell lines with many different cancer cell lines including drug resistant cell lines.

The cells can be shipped either as frozen or growing cultures, and are supplied complete with a specific data sheet together with a technical advisory pamphlet. Batch specific Certificates of Analysis and Authentication Records can be provided on request. ECACC can also supply cell lines on a standing order basis.

All cell lines added to the collection undergo full quality control and authentication procedures. These include testing for mycoplasma by a combination of culture isolation, Hoechst DNA staining and PCR, together with culture for containment bacteria, yeast and fungi.

Isoenzyme analysis is used to verify the species of each cell line deposit while DNA fingerprinting identifies cell line variation and/or cross contamination. ECACC routinely carries out two methods of DNA fingerprinting. Multilocus, applicable to most common species and STR DNA profiling for human cell lines only. The latter has the benefit of producing a digital 'fingerprint', which can be compared to ECACC’s extensive range of archived profiles. All HeLa contaminant cell lines are clearly identified in the full catalogue description.

Hybridoma Collection

This collection comprises over 400 monoclonal antibody-secreting hybridomas. The specification of the secreted antibodies covers a wide range of antigens including bacterial and viral; major histocompatability complex differentiation; oncogenes; plant and immunoglobulin.

ECACC can supply frozen or growing cultures of the hybridomas in the collection. In addition ECACC may also supply, assuming no restrictions are in place, purified antibody on a customized basis. The cost of providing purified antibody is dependent on the antibody production levels of the hybridoma, level of purification and quantity required.

All cell lines added to the collection undergo full quality control and authentication procedures. These include testing for mycoplasma by a combination of culture isolation, Hoechst DNA staining and PCR, together with culture for contaminant bacteria, yeast and fungi.

HLA Defined Human B-Lymphoblastoid Collection

This is a specialized collection of over 350 reference cell lines, originating from laboratories all over the world, to be used in the investigation of the HLA system. The collection was expanded in 1996 with cell lines from the 12th International Histocompatability Workshop (IHW) being added. A further 60 cell lines from the 13th IHW collection are due to be added later this year, along with a number of recently banked lines that were originally part of the 10th IHW collection.

Both serology and DNA typing data is available for the HLA Class I and II antigens together with information on non-HLA genes within the HLA region. This resource is available both as cell lines or DNA.

Human Random Control Collection

This is a collection of 500 lymphoblastoid cell lines derived from randomly selected Caucasian blood donors whose parents and grandparents were born in the UK or Ireland. DNA from these controls has already been extensively used in genome screening programs. This resource will be available as extracted DNA in a 96 well array format.

Full information on all the ECACC collections are available from the ECACC Website or in a CD-ROM format.

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