Sample Sets and Inventory Available:
Click the links below to download an inventory report spreadsheet. Please note that many of these inventory reports contain multiple tabs.
- Over 1,000 unique patient cases available. Most DTCs include cell population analysis and HLA-A02 typing by flow cytometry. Point mutation tested patient samples are listed on separate tab.
SCIENCE
Discovery scientists were the first to validate cryopreserved tissue dissociations as a viable alternative to sourcing fresh tissue for many research applications. With proprietary custom protocols, our scientists have consistently demonstrated high percentages of viable cells both pre-freeze (81.3%) and post-thaw (69.7%). We also have found that most cell types are largely unaffected by cryopreservation. Click to access the white paper confirming cell viability in Dissociate Tumor Cells (DTCs) via flow cytometry.
Our team conducted a flow cytometry analysis of over 400 tumor dissociations across 11 oncology indications to observe indication-specific trends in tumor compositions. The data from this large-scale flow cytometry analysis of tumor dissociations has been presented at scientific meetings.
Also, a Technical Note is now available that confirms the suitability of DTCs for downstream genomic analysis by whole exome and whole transcriptome sequencing.
We continually innovate the utility of tissue dissociations in contemporary research and downstream applications (Box 1). Leveraging sophisticated technologies and techniques, including flow cytometry and genomic applications, we are expanding our sample characterizations and laboratory services. Exploring these applications in R&D and through client partnerships, our team has cultivated a deep expertise in applying tissue dissociations to custom projects. We are available for consultations to optimize the results of studies involving our tissue dissociations.
SERVICE
Our tissue dissociations are delivered as cryopreserved vials of single cell suspensions with guaranteed cell counts following our Thawing Viable Cell Products protocol. Specimens are processed using validated procedures that have been optimized per disease indication. Each specimen is provided with the base clinical data from the patient case, along with a redacted pathology report.
Characterizations:
Our scientific team continually develops deeper characterization levels for our dissociated tissue inventories. View the characterization levels currently available in our existing inventories below:
- Base Patient Data – includes only base patient data and redacted pathology report
- Cell Populations – includes cell population analysis by flow cytometry + base patient data
- HLA-A02 Status – HLA-A02 status by flow cytometry + cell populations + base patient data
- Targeted Genomic Analysis – various biomarker and mutational statuses (MSI, EGFR, KRAS, etc.) by PCR or sequencing, without regard for characterization level
GENERAL INVENTORY: VIEW NOW
Box 1. Research Applications
- Flow Cytometry
- Sequencing – Technical Note
- Cell Isolation Studies
- Cell Culturing
- 3D Organoids
- Patient-Derived Xenografts (PDX)
- Identifying Novel Biomarkers
- Translational Development
- Monitoring Immune Activation
- Drug Response and Target Discovery
Frequently Asked Questions
How are dissociated tumor cells (DTCs) generated? What protocol is used?
Following surgical resection, tumors are placed in a proprietary tissue storage solution and shipped in a NanoCool™ Cooling System to Discovery’s expert Custom Biospecimen Processing Laboratory. Upon receipt, tumors are minced into small pieces and undergo a proprietary mechanical and enzymatic digestion to the single cell level. This digestion takes approximately 1 hour. Following digestion, cells are washed and cryopreserved.
Are red blood cells lysed prior to cryopreservation?
Red blood cell lysis is not performed.
Are dead cells or debris removed prior to cryopreservation?
Dead cells or debris removal is not performed, as previous attempts at these protocols have only provided modest reductions in the number of dead cells or amount of debris while negatively impacting cellular yields and composition.
How are DTCs cryopreserved?
DTCs are cryopreserved in CryoStor© CS10.
What are the recommended storage conditions for the cryopreserved DTCs?
DTCs are shipped on dry ice. Upon receipt, they should be used immediately or placed in liquid nitrogen vapor phase for long term storage.
How are the cell counts and viabilities of DTCs determined?
Cell counts and viability are determined using a Nexcelom Cellometer® with acridine orange and propidium iodide to identify live and dead nucleated cells, respectively, per the recommendations provided by Nexcelom. Given the cellular debris that can be present, we do not recommend the use of trypan blue-based cell counting methods, as this will overestimate the number of dead cells.
How is post thaw QC performed?
Following at least 24 hours in liquid nitrogen, one vial is removed from storage and quickly thawed in a 37°C water bath until only a small frozen crystal remains. Vials are transferred into a biosafety cabinet and diluted with 1 ml of DMEM/F12 + 10% FBS in a 15ml conical tube. 20µl of the cell suspension is mixed with 20µl of acridine orange/propidium iodide and counted on a Nexcelom Cellometer to determine cell counts and viability.
What cell surface markers are cleaved during the dissociation process?
We have profiled the sensitivity of numerous cell surface markers to the enzymatic cocktail used to dissociate tumors. For inquiries on specific markers, please contact info@dls.com.
My cell surface marker of interest hasn’t been tested for sensitivity. Can you test it?
Yes, we are able to test receptor sensitivity using samples with known expression, such as PBMCs or cell lines. For more information, please contact info@dls.com.
My cell surface marker is cleaved during the dissociation process. Are modified protocols available?
Yes, modified dissociation protocols are available that may maintain the expression of sensitive cell surface markers. While expression is preserved, the overall viability of the sample may be negatively impacted. For more information, please contact info@dls.com.
Do you have any recommendations for culturing DTCs?
For short term cultures using DTCs, we recommend using ultra-low attachment plates, such as Corning™ Costar™ Ultra-Low Attachment Microplates. Conventional tissue culture-treated plates should be avoided. Serum-free media is also recommended, and we highly recommend using penicillin, streptomycin, gentamicin, primocin, and amphotericin B to limit bacterial and fungal growth. For more information, please contact info@dls.com.
