Pseudo-Diaphragmatic Expansion Perfusion & Cryopreservation of Intact Viable Tissues


Researchers at MUSC have developed a rapid and reproducible method for the cryopreservation of a variety of tissues, such that when thawed are comparable to freshly resected tissues. To date, analysis has been completed on lung tissue from patients undergoing lobectomy or transplant and prostate tissue. This method involves the pseudo-diaphragmatic expansion (rapid and thorough perfusion) of pieces of freshly resected tissue utilizing a universal cryoprotectant (PDX-CP) followed by controlled-rate freezing in cryovials. Expansion-perfusion rates, volumes and the cryoprotectant formulation have been optimized for different clinical pathologies to maintain tissue architecture, decrease crystal formation and increase long-term cell viability. Morphology was greatly improved by the PDX-CP procedure compared with standard cryoprotectant formulations. Fresh versus post-thawed lung tissue showed minimal differences in histology, RNA integrity numbers and post-translational modified protein integrity (2-dimensional differential gel electrophoresis). It was possible to derive numerous live cell types, including alveolar epithelial cells, fibroblasts and stem cells, from the tissue for at least three months after cryopreservation. This new method will provide a uniform, cost-effective approach to the banking of biospecimens, with versatility to be amenable to any post-acquisition process applicable to fresh tissue samples.



Overview: With a CAGR of 7.9%, the global market value for biorepositories market is anticipated to be worth $23.9 billion by 2015, with North America holding approximately 30% of the market. Currently, the emphasis in biospecimen collection is standardization of pre-analytical variables. For example, there is an emphasis on noting and controlling the ischemic time for a tissue being harvested for preservation. An impressive list of factors (principally surgical and pathological) has been identified, and the hypothesis is that controlling and specifying these factors will enable collection of high quality biospecimens. However, it is also possible that even if all of these elements are controlled, we can still end up storing uniformly poor quality samples. The utility of human biospecimens as models is limited by specialized, post-acquisition tissue processing methods that preserve subclasses of analytes (e.g. RNA, protein, morphology) at the expense of others. A hallmark of these insufficiencies is the large strata of patients with diagnoses that are resistant to conventional treatments, indicative of patient subpopulations with alternate underlying mechanisms of disease or differing responses to treatments. Effective biospecimen collection and analysis hold the greatest promise for addressing these personalized medicine issues, permitting correlations between subpopulation responsiveness and associated molecular profile predictors. The biospecimens collected using current protocols fall short of their full potential utility for human disease research due to the variety of preservation protocols currently used that are analyte-dependent and not anticipatory of future novel analytical methodologies.


Applications: Biospecimen bank, therapeutic efficacy testing

Advantages:  Can utilize less samples with a more universal cryopreservation solution, one cryopreservation solution for multiple analytes and novel applications.

Key Words: Cryopreservation, lung, prostate, tissue, biospecimen, morphology, histology, pathology, ex vivo


Publications: BAATZ, JOHN E., et al. "Cryopreservation of Viable Human Lung Tissue for Versatile Post-thaw Analyses and Culture." In Vivo 28.4 (2014): 411-423.


Mancia, Annalaura, et al. "Cryopreservation and in vitro culture of primary cell types from lung tissue of a stranded pygmy sperm whale (Kogia breviceps)." Comparative Biochemistry and Physiology Part C: Toxicology & Pharmacology 155.1 (2012): 136-142.


Inventors: John E. Baatz & Demetri D. Spyropoulos

MUSC-FRD Technology ID: P1348

Patent Information:
For Information, Contact:
Docket BioPharma
Zucker Institute of Innovation Commercialization powered by MUSC
John Baatz
Demetri d. Spyropoulos
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