National Cancer Institute U.S. National Institutes of Health www.cancer.gov
Cooperative Human Tissue Network 1-866-462-2486

BIOSPECIMENS

SPECIMEN TYPES

The CHTN obtains tissues from routine surgical resections and autopsies. In addition, remnant body fluids (e.g., serum, leukapheresis products, pleural effusions) are available on some patients and limited histological services (e.g. frozen sections) can be obtained by special arrangement. Normal tissues (i.e. microscopically uninvolved by disease) commonly available from surgical resections include skeletal muscle, skin, bone, cartilage, connective tissue, breast, adipose tissue, small and large intestine, arteries, veins, thyroid, lung, gynecological tissues, kidney, spleen tonsils, testes, salivary glands and oral mucosa. Various pathological tissues also are obtained from surgical resections including head and neck malignancies and carcinomas of the lung, gastrointestinal tract, breast, genitourinary system, prostate, kidney and skin. In addition to the above more common specimens that are provided from surgical resections to investigators on a regional basis, access to rare tumors, e.g., neuroendocrine tumors and sarcomas, is provided from all divisions via networking. Pediatric tumors, diseased and normal tissues are available nationwide.

Additional normal tissues can be obtained from autopsies. These include liver, pancreas, adrenals, brain, pituitary, lymph nodes, heart and thymus.

BIOSPECIMEN COLLECTION

A detailed protocol for collection of biospecimens is developed in consultation with each investigator; however, the initial aspects of these protocols are usually similar. The usual protocol for collecting biospecimens from surgical resections is as follows:

  1. A list of biospecimens required by regional investigators and requests that have been networked are updated continuously.
  2. These biospecimen requests are matched with surgical resections so that each case provides maximal biospecimen resources to multiple investigators.
  3. When a potential research specimen is identified, the specimen is rapidly transferred from the operating room to a surgical pathologist (or pathology assistant) who provides CHTN personnel with remnant tissues that are not necessary for diagnosis.
  4. Similarly, autopsy biospecimens and remnant serum/plasma biospecimens not required for diagnosis are transferred by the pathology personnel to CHTN personnel.
  5. From this point, the biospecimens and fluids from both surgical resections and autopsies are processed according to the protocols of the individual researchers.

BIOSPECIMEN PROCESSING, PRESERVATION AND STORAGE

The processing of biospecimens varies according to the protocol of each individual investigator. Specimens can be snap-frozen in liquid nitrogen or frozen in cryoembedding media such as OCT and stored at liquid nitrogen or -80° C temperatures. Fresh tissue can be minced, placed in transfer media of choice or saline, and shipped on wet ice or at room temperature to the investigator for next day arrival. Similarly, for research involving substances stable after fixation, fixed or paraffin-processed biospecimens can be provided. In addition, some divisions offer limited histological processing by special arrangement. This includes stained or unstained paraffin embedded or frozen sections. Rare tumors and difficult to obtain specimens may be collected in excess of requests according to standard protocols adopted by the CHTN.

SHIPPING

The method of shipment of specimens varies with the type of tissue collected. Specimens stored at ultra-low temperatures may be batch shipped to investigators on dry ice. Fresh biospecimens are usually shipped out the day of collection on wet ice or at room temperature. Investigators can expect delivery of shipments by early the next morning.

TISSUE MICROARRAYS AND MACRODISSECTION

The Mid-Atlantic Division (http://faculty.virginia.edu/chtn-tma/home.html) currently has the following TMAs available to CHTN investigators. All unstained histologic sections are 4 microns thick on charged glass slides.

  • Normal Human Tissues TMA
  • Test TMA
  • Colorectal Carcinoma Progression TMA
  • Human Nervous System TMA
  • Breast Cancer Progression TMA
  • Ovarian Carcinoma Survey

The Pediatric Division houses the Children's Oncology Group (COG) Solid Tumor Tissue Bank, the COG Acute Lymphoblastic Leukemia bank and the Gynecologic Oncology Group (GOG) Tissue Bank. Both the COG and GOG have created tissue microarrays, which are available to approved researchers. Please contact Laura Monovich (614-722-2714 or Laura.Monovich@nationwidechildrens.org) for more information regarding these microarrays.

The CHTN also offers investigators macrodissected tissue. Macrodissection refers to gross manual dissection guided by analysis of a histologic section of a frozen tissue sample embedded in OCT, or of a fixed tissue sample embedded in paraffin, to create tissue samples enriched for a particular cell type. Two different methods of macrodissection are employed. The choice of method depends on the characteristics of the particular target tissue and the particular tissue sample to yield the greatest yield and enrichment of target cells, and on the requirements or preferences of the investigator to be served.

Though better defined than a non-dissected tissue sample, the actual percentage of targeted cells obtained by macrodissection can only be proscribed to the degree that tissue architecture and admixed cellular components allow. This will vary from tissue type to tissue type and from case to case. Although the percentage of target tissue can be estimated with a greater degree of precision than with a non-dissected tissue sample, this is still an estimate and not an absolute quantification of captured tissue.

QUALITY CONTROL

Quality control is fundamental to the successful operation of any service that provides tissue for research. For this reason, the Principal Investigators at each CHTN Division are experienced in anatomic pathology. The Principal Investigators, who are responsible for proper histopathological characterization, participate in research and understand the importance of quality control in the tissues provided.

All specimens are subject to an immediate gross examination by a pathologist. The diagnosis is then verified through frozen section, touch preparations, or subsequent evaluation of permanent histopathology. In most cases, the diagnosis can be assured before the specimen is released to the investigator. However, in some cases in which fresh specimens are needed, the tissues are provided to the investigator with a preliminary diagnosis, and the investigator is informed of the final diagnosis as soon as this information is available. In all divisions and institutions affiliated with the CHTN pathological diagnosis and patient care have total priority over the use of any specimen in research.

AT NO TIME WILL THE PATHOLOGICAL DIAGNOSIS OF A SPECIMEN BE COMPROMISED TO OBTAIN TISSUE FOR RESEARCH PURPOSES.

A feedback form or postcard is enclosed in each shipment, giving the investigator an opportunity to provide rapid information concerning the quality of that particular shipment. This information provides the divisions quick feedback and highlights any problems so they can be corrected immediately. A more detailed questionnaire is sent annually to investigators who receive tissue from the CHTN. The information collected in this annual evaluation will be used to identify positive features that should be maintained as well as problems that will be corrected.

BIOHAZARD AND INFECTION CONTROL

A major focus of the CHTN is preventing accidental exposure to infectious agents by research personnel who work with human tissues. Each investigator must agree to assume full responsibility for informing and training all personnel in the dangers of and procedures for safe handling of these and other human tissues. The CHTN has prepared a set of guidelines that provide detailed information and procedures for handling human tissues and body fluids used in research.1

Although this information is provided to CHTN investigators to assist them in developing working guidelines and operating procedures for their laboratory, each investigator remains responsible for the development and implementation of adequate safety procedures. This is formalized by an agreement on the application form that each investigator must sign.

In addition, the CHTN takes precautions to alert anyone opening a shipment that potentially biohazardous human tissue is enclosed. A biohazard label warning that the container holds human tissue and another label urging the reader to be familiar with the guidelines for handling human tissue before proceeding are placed on the inner packaging of each shipment.

The CHTN does not provide tissues from patients with known infections such as hepatitis, AIDS, or tuberculosis. However, because there is no universal policy on testing for infective agents such as hepatitis-B and HIV (AIDS virus) and because medical histories of patients are not always indicative of infection, the CHTN can never be absolutely sure that the tissues it provides are not biohazardous. For this reason, the CHTN requires its investigators to treat all specimens as if they are contaminated.

The CHTN will continue to evaluate and incorporate any changes in the recommended guidelines or development of new laboratory requirements for the safe handling of human tissues. The CHTN investigators will be informed of any new guidelines or developments.

1Grizzle WE, Bell W, Fredenburgh J. Safety in biomedical and other laboratories. In: Molecular Diagnostics. (Eds. G. Patrinos. W. Ansorg). Chapter 33, pp 421-428, 2005.

POLICY STATEMENT

The institution and Principal Investigators comprising the CHTN make every effort to ensure that there is an equitable distribution of tissue into and by the CHTN. In addition, several factors act to prevent CHTN tissue resources from benefiting only a limited number of investigators. Foremost, except for rare tumors, the number of surgical specimens is usually adequate for the recovery of research material.

BIOSPECIMEN PROCESSING FEES

Investigators pay a nominal processing fee for samples received from the CHTN. Slides and blocks to accompany frozen or fresh tissue specimens may be available for an additional fee. The charges are not for the tissue, but are to partially offset the costs of collecting, handling and preparing the specimens in accordance with the detailed requirements of the investigator. A sample is defined as one processed piece of a specimen, regardless of the sample size or type of processing. The investigator must also pay the cost of shipping samples to his/her laboratory. Click here for the CHTN processing fees in PDF.