Decrease in donor heart injury by recombinant clusterin protein in cold preservation with University of Wisconsin solution

doi:10.1016/j.surg.2011.09.032

Surgery

Volume 151, Issue 3, March 2012, Pages 364-371

Presented as a poster at American Transplant Congress in Philadephia, PA, April 30–May 4, 2011.

https://doi.org/10.1016/j.surg.2011.09.032 Get rights and content

Background

Donor organ injury during cold preservation before transplantation negatively impacts graft survival. Clusterin (CLU) is a chaperonic protein, and its expression confers donor hearts resistance to cold ischemic injury. This study was conducted to evaluate if the supplement of recombinant CLU protein (rCLU) protects donor organs from injury during cold storage with University of Wisconsin (UW) solution.

Methods

Human endothelial cell cultures were used as an in vitro model. Heart transplantation in mice was used as an in vivo model. Cell membrane disruption or death was indicated by the release of lactate dehydrogenase (LDH). Donor injury was determined by its functional recovery, and histologic and biochemical analyses.

Results

Supplement of rCLU to UW solution protected cultured human endothelial cells from cold-induced cell necrosis, as evidenced by a decrease in both release of LDH and the number of ethidium bromide-stained necrotic cells. The protective activity of rCLU was associated with enhanced membrane fluidity at cold temperature. During cold storage of heart organs in UW solution, supplemental rCLU significantly decreased LDH release from heart tissue. In a preclinical model of transplantation, heart grafts after cold preservation with rCLU-containing UW solution had better functional recovery and decreased perivascular inflammation, neutrophil infiltration, and cardiac cell death, including apoptosis and necrosis, that correlated with lower levels of serum creatine kinase and LDH in recipients.

Conclusion

Our data suggest that supplement of CLU protein in a cold preservation solution may have potential in improving cold preservation of donor organs in transplantation.

Section snippets

Animals and cell culture

C57BL/6j (B6) mice (males, 8–10 weeks old) were purchased from Jackson Laboratory (Bar Harbor, ME). All animal use procedures in this study were performed and monitored in accordance with the Canadian Council on Animal Care guideline under the protocols approved by the Animal Use Subcommittee at the University of British Columbia.

Primary human umbilical vein endothelial cells (HUVECs) were purchased from Lonza Walkersville (Walkersville, MD) and were immortalized with origin-deficient SV40 DNA

Preventing cell membrane disruption or necrosis in cultured HUVECs at cold temperature

When the plasma membrane was disrupted or lysed (necrosis), cytoplasmic LDH was released to the medium and its level was used as a biomarker for cell lysis or necrosis. The addition of rCLU to UW solution (Viaspan; Duramed Pharm Inc, Pomona, NY) prevented the cell lysis of cultured HUVECs in a dose-dependent manner, as evidenced by a significant decrease in LDH release from 73 ± 7% in HUVECs treated with UW solution alone (control) to 26 ± 8% in those with 0.5 mg/mL of rCLU or to 23 ± 2% with 1

Discussion

Cold ischemic injury, together with additional injury from rewarming or reperfusion, contributes to poor organ function in the immediate posttransplantation and subsequent rejection episodes.18, 19, 20 Therefore, the limitation of cold ischemic injury by optimizing cold preservation may improve the success of organ transplantation. Our previous study found that graft-expressing CLU renders resistance of donor hearts to cold ischemic injury in an experimental study.¹⁶ The present study shows

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Citation Excerpt :
When Clusterin (CLU) a chaperonic protein was added, the results showed a reduction in donor heart injury during preservation at 4 °C before transplantation (Wyatt et al., 2009). Therefore, the presence of CLU added to the UW solution gives donor hearts greater resistance to cold ischemic injury (Guan et al., 2012). However, conventional refrigeration and freezing have their limitations, since they require storage at low temperatures and special transport conditions, as well as competent personnel (Taylor et al., 2004).
With the advancement of science and technology, it is crucial to have effective preservation methods for the stable long-term storage of biological material (biomaterials). As an alternative to cryopreservation, various techniques have been developed, which are based on the survival mechanism of anhydrobiotic organisms. In this sense, it has been found that the synthesis of xeroprotectants can effectively stabilize biomaterials in a dry state. The most widely studied xeroprotectant is trehalose, which has excellent properties for the stabilization of certain proteins, bacteria, and biological membranes. There have also been attempts to apply trehalose to the stabilization of eukaryotic cells but without conclusive results. Consequently, a xeroprotectant or method that is useful for the stable drying of a particular biomaterial might not necessarily be suitable for another one. This article provides an overview of recent advances in the use of new techniques to stabilize biomaterials and compare xeroprotectants with other more standard methods.
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: Supported by the Centres of Excellence for Commercialization and Research and Natural Sciences and Engineering Council of Canada. Dr Li received the Postdoctoral Training Award from the Canadian Institutes of Health Research Training Program in Transplantation (Vancouver Coastal Health Research Institute).

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Original CommunicationDecrease in donor heart injury by recombinant clusterin protein in cold preservation with University of Wisconsin solution

Background

Methods

Results

Conclusion

Section snippets

Animals and cell culture

Preventing cell membrane disruption or necrosis in cultured HUVECs at cold temperature

Discussion

Transpl Proc

Transplant Proc

Transplant Proc

Lancet

Int J Biochem Cell Biol

J Biol Chem

J Heart Lung Transplant

Cryobiology

Theriogenology

Cryobiology

J Biol Chem

Kidney Int

J Thorac Cardiovasc Surg

Organ preservation

Annu Rev Med

Organ preservation solutions for whole organ pancreas transplantation

Curr Opin Organ Transplant

Organ reperfusion and preservation

Front Biosci

Perspectives in organ preservation

Transplantation

Original Communication
Decrease in donor heart injury by recombinant clusterin protein in cold preservation with University of Wisconsin solution