Biobanking and Biospecimen Quality - Inseparable Entities

It is critical to determine the best method for storing tissue. When researchers request tissue from our biobank, we must ensure that we are providing them with the highest quality tissue possible. Therefore, the biobank at Windber Research Institute (WRI) is constantly designing and performing experiments to determine how the tissue collected from the many donors is impacted by pre-analytical variables and how these “uncontrollable” factors could impact the derivatives of the tissue such as DNA, RNA and Protein.

Pre analytical variables may be represented by the following 1) Physiology of the human research participant 2) Specimen collection practices 3) Specimen handling practices before downstream testing (NCI Best practices, 2014; http://biospecimens.cancer.gov/bestpractices/to/bcpsrd.asp).  There are other variables that could potentially affect tissue quality, for example,  it is believed that “minor” aspects such as sample storage vessels (snap cap and screw lid, non-sterile, sterile or RNase/DNase free tubes) could impact tissue quality (Pfaendner 2006). Additionally, tubes can be made out of different plastics which can result in adherence of biomolecules to the vessel walls or leaching of plastic components into the sample (McDonald et al., 2008). This makes it important to ensure that samples are stored in appropriate tubes that will maintain tissue quality over time and at specific storage conditions (Smith 2011). Additionally, the storage temperature of tissue and nucleic acids is very critical (Andreasson et al. 2013). Normally, samples intended for long term storage are stored in liquid nitrogen vapor tanks at -190 degrees Celsius and samples intended for short term storage are stored in -80 degree Celsius freezers (Karisson and Toner 1996). However, not all biobanks have the ability to store in liquid nitrogen vapor and as such utilize mechanical freezers at -80 degree Celsius.  It is essential for biobankers to know which types of samples can be stored at which temperature and for what length of time (Hubel, Spindler and Skubitz 2014). These are the many areas awaiting evidence based data to support biobank storage protocols. Biospecimen Science Research is the umbrella whereby these issues are being addressed and in the future, this science should provide more evidence based data to allow biobanks fully understand all that impacts tissue quality.  Since biobanks have their individual interest in what they store, what these tissue samples are used for and for how long they will be stored, these evidence based studies will have to meet a variety of needs.

Interestingly, studies have shown that the concentration of the RNA stored could have an impact on the integrity of the RNA over time (Olivieri et al. 2014). In their study Olivieri et. al, showed the effects of storing RNA at high concentrations (250ng/µL) and low concentration (25ng/µL). They found that after 8 months of storage, RNA stored at high concentrations preserved its integrity while RNA stored at low concentrations showed significant degradation. Therefore, while low concentrations of RNA may be acceptable for immediate analysis, storage for a few months may not be suitable. These kinds of results are a clear indication of the complex issues that biobanks face when working to maintain tissue integrity and provide quality DNA, RNA and Protein for downstream analysis.

Proper storage of tissue and nucleic acids can greatly affect experiments and their outcome (Yang and Chang 2012). If tissue is not stored correctly, valuable proteins can be degraded (Auer et al. 2014). Additionally, degradation of RNA and/or DNA can affect critical analyses such as gene expression (Shane, Kohlmeyer & Hunter 2010). Biobanks need to be on top of these issues either by designing internal experiments that provide data for the design of evidence based Standard Operating Procedures (SOPs) or keeping abreast with research results in the area of Biospecimen Science as the source of new information/findings that will drive biobanking SOPs. The International Society for Biological and Environmental Repositories (ISBER  http://www.isber.org/) and the Biorepository and Biospecimen Research Branch of the NCI (BBRB  http://biospecimens.cancer.gov/default.asp) remain great resources for biobanks to tap into for current ideas and information that will enable them maintain the desired standards that will drive biobanking for quality tissue. 

References

Andreasson, A., Kiss, NB., Juhlin, CC and Höög, A (2013). Long-Term Storage of Endocrine Tissues at -80^oC Does Not Adversely Affect RNA Quality or Overall Histomorphology. Biopres. & Biobanking, 10.1089/bio.2013.0038

Auer, H., Mobley, J., Ayers, L., Bowen, J., Chuaqui, R., Johnson, L., … Ramirez, N. (2014). The effects of frozen tissue storage conditions on the integrity of RNA and protein. Biotechnic & Histochemistry : Official Publication of the Biological Stain Commission, 89(7), 518–528. http://doi.org/10.3109/10520295.2014.904927

Hubel, A., Spindler, A and Skubitz, A (2014). Storage of Human Biospecimens: Selection of the Optimal Storage Temperature. Biopres. & Biobanking, 10.1089/bio.2013.0084

Karlsson, JOM and Toner, M (1996). Long-term storage of tissues by cryopreservation: critical issues. Biomaterials, 17(3): 243-256.

McDonald, GR et al., (2008). Bioactive contaminants leach from disposable laboratory plasticware.  Science, 322(5903):917

Olivieri, EHR., Franco, LA., Pereira, RG., Mota, LDC., Campos, AH and Carraro, DM (2014). Biobanking Practice: RNA Storage at Low Concentrations Affects Integrity. Biopres. & Biobanking, 10.1089/bio.2013.0056

Pfaendner, R (2006). How will additives shape the future of plastics? Polymer Degrad. Stabil. 91:2249-2256.

Shane, M. L., Kohlmeyer, M., Hunter, T., and Tighe, S. (2010). RT-qPCR Analysis of Degraded RNA using Five Different Pre-Amplification Methods. Journal of Biomolecular Techniques : JBT, 21(3 Suppl), S45.

Smith, C (2011). Selecting the Right Tubes or Plates for Your Research. Life Science Articles, http://www.biocompare.com

Yang, T-H and Chang, P-L (2012). Determination of RNA degradation by capillary electrophoresis with cyan light-emitted diode-influences fluorescence. J. of Chromatogr. A, http://dx.doi.org/10.1016/j.chroma.2012.03.070

THE BIOREPOSITORY AT WINDBER RESEARCH INSTITUTE (WRI) RECEIVES CAP ACCREDITATION

Biobanking is the fastest growing component in translational research and the Windber Research Institute established a biorepository in 2001 with the mission of “Providing high quality biospecimens to enhance the scope and reliability of molecular research.”

We are happy to announce that the Accreditation Committee of the College of American pathologists (CAP) has awarded accreditation to the WRI Biorepository based on the results of an onsite inspection on 4/7/2015.

The Senior Director of the Biorepository, Dr. Stella Somiari, PhD, was advised of this national recognition on 5/14/2015 and congratulated for the excellence of the services being provided. The repository at WRI is one of the 39 CAP accredited biorepositories to date and one of the 2 CAP accredited biorepositories in Pennsylvania!

The accreditation program for biorepositories by CAP was started in 2012 and is the first of such programs offered for biorepositories.  It is a three-year accreditation cycle which includes on-site inspection, desk review, and optional education modules and gap assessment. As part of the on-site inspection, the CAP uses Accreditation Checklists to provide a comprehensive and up-to-date blueprint of quality practices to assist biorepositories in improving their operations and ensure quality. Further, a desk review offers a remote review of a biorepository’s quality plan, certain procedures and select quality and process statistics. The program is designed to ensure that standards are maintained and there is uniformity in all operational procedures which should result in the management and distribution of the highest quality biospecimens.   

The recent CAP accreditation status for the WRI puts the official “stamp” of quality to all its operations and with the added oversight by CAP users of our biorepository resources will have increased confidence in the quality of the biospecimens which we provide to support high impact molecular research.  We are proud to continue to serve the John P. Murtha Cancer Center  of Walter Reed Bethesda in the many capacities that we do, even more so, as an accreditaed biorepository.

About the WRI Biorepository

The biorepository at WRI was established in 2001 to manage specimen procurement, processing, storage, and distribution to the Department of Defense funded Clinical Breast Care Project (CBCP). To date, its activities have expanded to include biobanking activities for the John P. Murtha Cancer Center of Walter Reed, Bethesda, MD.  The repository has established and maintained extensive Quality Management Systems (QMS) to cover all areas of its operations and participates in regular Proficiency/Quality Testing activities in areas that include DNA/RNA Extraction and Quantification, RNA Integrity assessment, and Tissue Histology. The repository has expertise in the establishment and management of multiple collection sites. It has experience working with regional, urban, and community hospitals and has managed collections from international sites and organ procurement organizations. Tissue processing is performed in a variety of formats and new processing protocols are developed as needed. Services provided include:

General

Acquisition, processing and distribution of human biospecimens for research

Current inventory available for research includes serum, plasma, DNA, RNA, PAXgene Blood (for RNA extraction), assorted tissue types (breast, heart, kidney, lung, colon, and lymph node, etc)

Laser Microdissection (LMD) of Tissue for Research

General Histology Services

Long or Short Term Biospecimen Storage in ultra-low or liquid nitrogen freezers

DNA and RNA Extraction

Nucleic Acid Extraction from a variety of biological materials

Nucleic Acid Quality Assessment using a variety of technologies

Consulting Services

Customized Biorepository Design and Setup.

Development of Standard Operating Procedures (SOP) for biorepositories.

Design and Review of Biospecimen Research Study Protocols.

In recent years, Biospecimen Science Research has become an integral part of the biorepository’s operations. Current biospecimen research activities include elucidation of the effects of pre- analytical variables on biospecimen quality, effects of storage time and condition of storage on biospecimen quality, evaluation of new collection methods that will not only conserve scare tissue but also allow the banking of minimal tissue from surgical procedures which would normally have not been available for research and banking.  The repository is also conducting cutting-edge research aimed at developing non-destructive methods for tissue quality assessment.