The Leukemia Tissue Bank Shared Resource (HMSR) facilitates successful translation of basic research to the clinical setting by providing an extensive repository of tissue samples from patients with leukemia, lymphoma or myeloma, accompanying pathologic, cytogenetic and clinical data for correlation of clinical and biological results, and expert training and support for cancer investigators.
An NCI-sponsored biorepository, the LTBSR has more than 40,000 vials of cryopreserved viable cells and 13,000 vials of matched frozen plasma and/or serum samples from more than 4,000 patients treated for leukemia and other hematologic malignancies.
The LTBSR’s services include:
Tumor Cell Isolation, Derivative Sample Preparation and Cryopreservation of Viable Cells
Cells from bone marrow aspirates and peripheral blood samples are isolated by Ficoll density gradient centrifugation. After a viable cell count is completed, cells are washed and allocated for derivative sample preparation as determined by the corresponding treatment, correlative studies or research protocol. Remaining viable cells are cryopreserved and stored long-term in a liquid nitrogen freezer.
Cytological Slide Preparation
The cytological centrifuge is a specialized instrument to deposit cells uniformly into a defined area on a glass slide. These slides are then preserved and stored for future microscopic evaluations (e.g., immunocytochemistry).
Dangerous Goods Shipping
Packages containing human samples and/or dry ice require special shipping procedures. The LTBSR is experienced in the proper routes to ensure safe delivery of samples to off-campus recipients.
DNA Isolation from Mononuclear Cells
To investigate genetic/epigenetic changes that may occur with disease, the LTBSR isolates high-quality DNA from fresh or cryopreserved mononuclear cell fractions of bone marrow aspirate or peripheral blood samples.
DNA Isolation from Buccal Swabs
DNA isolation from buccal (cheek lining) cells collected on a swab or in mouthwash provides normal germline DNA from our patients so that disease-specific changes can be identified.
Materials Processing for Studies
The LTBSR can process material for studies including protein, RNA and DNA, and also can prepare cDNA from RNA samples. The resource also coordinates central molecular and flow cytometry analyses for studies as needed. Central isolation of protein and nucleic acids has allowed the LTBSR to provide investigators with the exact amount of uniform, high-quality material required, thereby conserving material obtained from a single vial of cells for additional studies. We make every attempt to customize sample handling, processing and storage to suit each clinical trial we support.
PAXgene Sample Processing
In cases where immediate Ficoll separation is not feasible, the LTBSR utilizes t he PAXgene Blood system. This consists of specialized tubes for blood collection and stabilization as well as purification of high-quality DNA or RNA in a single-tube procedure.
Primary Cell Culture
Following isolation from blood or marrow, patient tumor cells can be cultured for a limited time in sterile media. This procedure is useful for testing experimental agents or conditions, allowing the researcher to monitor changes in live tumor cells under carefully controlled conditions.
Quality Control of Derivative Specimens
This includes highly sensitive analysis of DNA, RNA and protein preparations via a new Qubit fluorometer. This instrument enables quick determination of concentration and quality of isolates while preserving the bulk of the sample for research.
Research Consultation – Sample Selection and Data Processing
Our staff will work with investigators to customize sample procurement, handling, processing and storage. We work with investigators to identify samples in the biorepository to match their research requirements.
RNA Isolation from Mononuclear Cells
We prepare RNA according to a variety of methodologies to best suit requirements of the individual research project.
Sample Retrieval, Packaging and Shipping
Once a sample set has been identified for a specific clinical trial or research study, we retrieve samples from long-term storage (and, if needed, prepare derivative specimens) to be released either to investigators at Ohio State or collaborators around the world.
Serum or Plasma Isolation
Serum (fluid from clotted blood) and plasma (fluid from non-clotted blood) can be derived from whole blood as needed according to the research protocol.
Wright/Giemsa with Blast Count
The LTBSR can perform staining and microscopic evaluation of cytological slides to determine percent of tumor cells in a sample.
The LTBSR operates in accordance with NCR Best Practices for sponsored repositories, and offers OSUCCC members and the cancer research community access to the following instrumentation, equipment and supplies:
Beckman Coulter ViCell ™ Counter for cell counts and viability data on each Ficoll-separated blood or marrow sample prior to cryopreservation.
Benchtop Centrifuges for mononuclear cell separation
Biological safety Cabinets for sterile processing (and if needed, cell culture) of samples
Buccal Swab and Mouthwash Collection Kit s for the collection and extraction of normal DNA
Computer Workstations for electronic database management
Dangerous Goods Shipping Containers for sending samples to approved outside investigators
Dry Ice for on-campus transport or shipping
MVE Automated Filling System for efficient liquid nitrogen (LN2) use. The system includes a service area and automatic tank switcher where up to four LN2 tanks can be stationed. Vacuum-jacketed piping leads to filling stations for each LN2 freezer.
MVE LN2 Freezers for safe long-term sample storage without direct contact with LN2. Freezers hold 15,500-36,400 samples, depending on rack size and storage box profile.
NanoDrop Spectrophotometer for spectrophotometric analysis of absorbance ratios to quantify derivative specimens. This spectrophotometer quickly determines the concentration of RNA, DNA or protein, and uses an extremely small sample volume. This enables the LTBSR to perform appropriate quality checks of samples while preserving the bulk of the sample for research.
Polymerase Chain Reaction Thermocycler for quality control of procured and prepared materials
Rees Scientific Environmental Monitoring System for 24/7 monitoring of all LTBSR freezers and refrigerators. The Rees system provides an hourly readout of monitored units and immediate telecommunication with individuals in the event of temperature fluctuation beyond the specified range.
Refrigerated Microcentrifuges for derivative sample preparation
QIAcube™ Biorobot for automated sample preparation for RNA, DNA and protein using Qiagen® spin-column technology. The QIAcube automates simultaneous sample preparation for more consistent sample products and improved lab efficiency.
Qubit Fluorometer for highly sensitive determination of the concentration and quality of derivative specimens while preserving the bulk of the sample for research.
Buccal swab DNA collection kit – CCC Member
Dangerous goods shipping charge – CCC Member
Dangerous goods shipping container – CCC Member
DNA isolation from buccal swab – CCC Member
Dry ice (for transport/shipping) – CCC Member
Positive selection/depletion (1st sample)
Positive selection/depletion (each add. 10 samples)
Preparation of cytological slides –- CCC Member
Procure w/RNA or DNA lysis prep only – CCC Member
Sample prep-lymph node/small tissue biopsy – CCC Member
Sample retrieval (per 10 samples) – CCC Member
Search charge – Bucci – CCC Member
Search charge – Faculty – CCC Member
Serum/plasma prep w/cryopres – CCC Member
Standard PBMC procure w/cryopres and LN2 (<10vials)
Technician time – CCC Member
Wright/Giemsa with blast count – CCC Member
DNA isolation (thawing cells required)
Thaw viable cells for RNA or DNA isolation
BD Cell Prep Tube (mononuclear cells)
RNA isolation – Qiagen (thawing cells required)
RNA isolation – Trizol (thawing cells required)