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Research ArticleExperimental Studies

Cryopreservation of Viable Human Lung Tissue for Versatile Post-thaw Analyses and Culture

JOHN E. BAATZ, DANFORTH A. NEWTON, ELLEN C. RIEMER, CHADRICK E. DENLINGER, E. ELLEN JONES, RICHARD R. DRAKE and DEMETRI D. SPYROPOULOS
In Vivo July 2014, 28 (4) 411-423;
JOHN E. BAATZ
1Department of Pediatrics, Medical University of South Carolina, Charleston, SC, U.S.A.
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  • For correspondence: baatzje@musc.edu
DANFORTH A. NEWTON
1Department of Pediatrics, Medical University of South Carolina, Charleston, SC, U.S.A.
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ELLEN C. RIEMER
2Department of Pathology and Laboratory Medicine, Medical University of South Carolina, Charleston, SC, U.S.A.
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CHADRICK E. DENLINGER
3Department of Surgery, Medical University of South Carolina, Charleston, SC, U.S.A.
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E. ELLEN JONES
4Department of Pharmacology, Medical University of South Carolina, Charleston, SC, U.S.A.
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RICHARD R. DRAKE
4Department of Pharmacology, Medical University of South Carolina, Charleston, SC, U.S.A.
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DEMETRI D. SPYROPOULOS
2Department of Pathology and Laboratory Medicine, Medical University of South Carolina, Charleston, SC, U.S.A.
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Abstract

Clinical trials are currently used to test therapeutic efficacies for lung cancer, infections and diseases. Animal models are also used as surrogates for human disease. Both approaches are expensive and time-consuming. The utility of human biospecimens as models is limited by specialized tissue processing methods that preserve subclasses of analytes (e.g. RNA, protein, morphology) at the expense of others. We present a rapid and reproducible method for the cryopreservation of viable lung tissue from patients undergoing lobectomy or transplant. This method involves the pseudo-diaphragmatic expansion of pieces of fresh lung tissue with cryoprotectant formulation (pseudo-diaphragmatic expansion-cryoprotectant perfusion or PDX-CP) followed by controlled-rate freezing in cryovials. Expansion-perfusion rates, volumes and cryoprotectant formulation were optimized to maintain tissue architecture, decrease crystal formation and increase long-term cell viability. Rates of expansion of 4 cc/min or less and volumes ranging from 0.8-1.2 × tissue volume were well-tolerated by lung tissue obtained from patients with chronic obstructive pulmonary disease or idiopathic pulmonary fibrosis, showing minimal differences compared to standard histopathology. Morphology was greatly improved by the PDX-CP procedure compared to simple fixation. 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 cell types, including alveolar epithelial cells, fibroblasts and stem cells, from the tissue for at least three months after cryopreservation. This new method should 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.

  • Cryopreservation
  • lung
  • biorepository
  • biobank
  • Received March 10, 2014.
  • Revision received March 18, 2014.
  • Accepted March 20, 2014.
  • Copyright © 2014 International Institute of Anticancer Research (Dr. John G. Delinassios), All rights reserved
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Vol. 28, Issue 4
July-August 2014
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Cryopreservation of Viable Human Lung Tissue for Versatile Post-thaw Analyses and Culture
JOHN E. BAATZ, DANFORTH A. NEWTON, ELLEN C. RIEMER, CHADRICK E. DENLINGER, E. ELLEN JONES, RICHARD R. DRAKE, DEMETRI D. SPYROPOULOS
In Vivo Jul 2014, 28 (4) 411-423;

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Cryopreservation of Viable Human Lung Tissue for Versatile Post-thaw Analyses and Culture
JOHN E. BAATZ, DANFORTH A. NEWTON, ELLEN C. RIEMER, CHADRICK E. DENLINGER, E. ELLEN JONES, RICHARD R. DRAKE, DEMETRI D. SPYROPOULOS
In Vivo Jul 2014, 28 (4) 411-423;
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Keywords

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