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5.1
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Cells
Special Issues
Biobanking of Engineered and Natural Tissues
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Biobanking of Engineered and Natural Tissues

A special issue of Cells (ISSN 2073-4409). This special issue belongs to the section "Cellular Biophysics".

Deadline for manuscript submissions: closed (31 May 2022) | Viewed by 20688

Special Issue Editors

Dr. Kelvin G. M. Brockbank

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Guest Editor
Clemson University, Clemson, SC, USA
Tissue Testing Technologies LLC, North Charleston, SC, USA
Interests: biomaterials preservation; tissue engineering; regenerative medicine
Dr. Lia H. Campbell

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Guest Editor
Tissue Testing Technologies LLC, North Charleston, SC, USA
Interests: biomaterials preservation; tissue engineering; regenerative medicine

Special Issue Information

Dear Colleagues,

There are huge potential markets for research on the diagnostic and transplantation applications of cells, tissues, and organs. A strategic assessment of the field from the Multi-Agency Tissue Engineering Science (MATES) group of federal agencies provided eight critical priorities for the field, three of which relate directly to the need for better preservation methods. Another more recent analysis of strategic directions in tissue engineering also highlighted preservation issues. Anecdotally, we have heard that another more recent MATES review has given preservation methods an even higher priority for regenerative medicine. Biobanking has the potential to save and improve many millions of lives. The last decade has seen tremendous proof-of-principle advances such as the cryopreservation of sheep ovaries, human fingers, as well as the successful storage of mammalian livers at high subzero temperatures. In this Special Issue we are seeking manuscripts on work focused on novel and innovative strategies to improve the quality of banked tissues, which may include strategies such as, but not limited to, ice-free vitrification, nanowarming, high subzero storage, nature-inspired cryotolerance, hypothermic perfusion, multi-thermic perfusion, computational modeling, ice control, isochoric cryopreservation, and methods for assessing biobanking outcomes (e.g., exciting new imaging methods). This Special Issue of Cells should improve our understanding of the potential impact of novel biobanking strategies in development, identifying where these new methods work and their current limitations. This Issue should include researchers working on autologous, allogeneic, and tissue-engineered tissue models and therapeutic products.

Dr. Kelvin G. M. Brockbank
Dr. Lia H. Campbell
Guest Editors

Manuscript Submission Information

Manuscripts should be submitted online at www.mdpi.com by registering and logging in to this website. Once you are registered, click here to go to the submission form. Manuscripts can be submitted until the deadline. All submissions that pass pre-check are peer-reviewed. Accepted papers will be published continuously in the journal (as soon as accepted) and will be listed together on the special issue website. Research articles, review articles as well as short communications are invited. For planned papers, a title and short abstract (about 100 words) can be sent to the Editorial Office for announcement on this website.

Submitted manuscripts should not have been published previously, nor be under consideration for publication elsewhere (except conference proceedings papers). All manuscripts are thoroughly refereed through a single-blind peer-review process. A guide for authors and other relevant information for submission of manuscripts is available on the Instructions for Authors page. Cells is an international peer-reviewed open access semimonthly journal published by MDPI.

Please visit the Instructions for Authors page before submitting a manuscript. The Article Processing Charge (APC) for publication in this open access journal is 2700 CHF (Swiss Francs). Submitted papers should be well formatted and use good English. Authors may use MDPI's English editing service prior to publication or during author revisions.

Keywords

  • Hypothermic storage
  • normothermic storage
  • perfusion
  • cryopreservation
  • vitrification
  • freezing
  • cold chain
  • nanowarming
  • transplantation
  • tissue engineering
  • regenerative medicine

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Published Papers (6 papers)

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Research

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16 pages, 6131 KiB  
Article
Epigenetic Alterations in Cryopreserved Human Spermatozoa: Suspected Potential Functional Defects
by Wanxue Wang, Plamen Todorov, Cheng Pei, Mengying Wang, Evgenia Isachenko, Gohar Rahimi, Peter Mallmann and Vladimir Isachenko
Cells 2022, 11(13), 2110; https://doi.org/10.3390/cells11132110 - 4 Jul 2022
Cited by 5 | Viewed by 2809
Abstract
Background: Gene set enrichment analysis (GSEA) was conducted on raw data, and alternative splicing (AS) events were found after mRNA sequencing of human spermatozoa. In this study, we aimed to compare unknown micro-epigenetics alternations in fresh and cryopreserved spermatozoa to evaluate the effectivity [...] Read more.
Background: Gene set enrichment analysis (GSEA) was conducted on raw data, and alternative splicing (AS) events were found after mRNA sequencing of human spermatozoa. In this study, we aimed to compare unknown micro-epigenetics alternations in fresh and cryopreserved spermatozoa to evaluate the effectivity of cryopreservation protocols. Methods: Spermatozoa were divided into three groups: fresh spermatozoa (group 1), cryoprotectant-free vitrified spermatozoa (group 2), and conventionally frozen spermatozoa (group 3). Nine RNA samples (three replicates in each group) were detected and were used for library preparation with an Illumina compatible kit and sequencing by the Illumina platform. Results: Three Gene Ontology (GO) terms were found to be enriched in vitrified spermatozoa compared with fresh spermatozoa: mitochondrial tRNA aminoacylation, ATP-dependent microtubule motor activity, and male meiotic nuclear division. In alternative splicing analysis, a number of unknown AS events were found, including functional gene exon skipping (SE), alternative 5′ splice sites (A5SS), alternative 3′ splice sites (A3SS), mutually exclusive exon (MXE), and retained intron (RI). Conclusions: Cryopreservation of spermatozoa from some patients can agitate epigenetic instability, including increased alternative splicing events and changes in crucial mitochondrial functional activities. For fertilization of oocytes, for such patients, it is recommended to use fresh spermatozoa whenever possible; cryopreservation of sperm is recommended to be used only in uncontested situations. Full article
(This article belongs to the Special Issue Biobanking of Engineered and Natural Tissues)
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15 pages, 2526 KiB  
Article
Ice Control during Cryopreservation of Heart Valves and Maintenance of Post-Warming Cell Viability
by Kelvin G. M. Brockbank, John C. Bischof, Zhenzhen Chen, Elizabeth D. Greene, Zhe Gao and Lia H. Campbell
Cells 2022, 11(12), 1856; https://doi.org/10.3390/cells11121856 - 7 Jun 2022
Cited by 8 | Viewed by 2875
Abstract
Heart valve cryopreservation was employed as a model for the development of complex tissue preservation methods based upon vitrification and nanowarming. Porcine heart valves were loaded with cryoprotectant formulations step wise and vitrified in 1–30 mL cryoprotectant formulations ± Fe nanoparticles ± 0.6 [...] Read more.
Heart valve cryopreservation was employed as a model for the development of complex tissue preservation methods based upon vitrification and nanowarming. Porcine heart valves were loaded with cryoprotectant formulations step wise and vitrified in 1–30 mL cryoprotectant formulations ± Fe nanoparticles ± 0.6 M disaccharides, cooled to −100 °C, and stored at −135 °C. Nanowarming was performed in a single ~100 s step by inductive heating within a magnetic field. Controls consisted of fresh and convection-warmed vitrified heart valves without nanoparticles. After washing, cell viability was assessed by metabolic assay. The nanowarmed leaflets were well preserved, with a viability similar to untreated fresh leaflets over several days post warming. The convection-warmed leaflet viability was not significantly different than that of the nanowarmed leaflets immediately after rewarming; however, a significantly higher nanowarmed leaflet viability (p < 0.05) was observed over time in vitro. In contrast, the associated artery and fibrous cardiac muscle were at best 75% viable, and viability decreased over time in vitro. Supplementation of lower concentration cryoprotectant formulations with disaccharides promoted viability. Thicker tissues benefited from longer-duration cryoprotectant loading steps. The best outcomes included a post-warming incubation step with α-tocopherol and an apoptosis inhibitor, Q-VD-OPH. This work demonstrates progress in the control of ice formation and cytotoxicity hurdles for the preservation of complex tissues. Full article
(This article belongs to the Special Issue Biobanking of Engineered and Natural Tissues)
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14 pages, 5671 KiB  
Article
Gluconate-Lactobionate-Dextran Perfusion Solutions Attenuate Ischemic Injury and Improve Function in a Murine Cardiac Transplant Model
by Yinan Guo, Franka Messner, Sarah E. Beck, Marcos Iglesias Lozano, Hubert Schwelberger, Yichuan Zhang, Kai Kammers, Byoung Chol Oh, Elizabeth D. Greene, Gerald Brandacher and Kelvin G. M. Brockbank
Cells 2022, 11(10), 1653; https://doi.org/10.3390/cells11101653 - 16 May 2022
Cited by 3 | Viewed by 2310
Abstract
Static cold storage is the cheapest and easiest method and current gold standard to store and preserve donor organs. This study aimed to compare the preservative capacity of gluconate-lactobionate-dextran (Unisol) solutions to histidine-tryptophan-ketoglutarate (HTK) solution. Murine syngeneic heterotopic heart transplantations (Balb/c-Balb/c) were carried [...] Read more.
Static cold storage is the cheapest and easiest method and current gold standard to store and preserve donor organs. This study aimed to compare the preservative capacity of gluconate-lactobionate-dextran (Unisol) solutions to histidine-tryptophan-ketoglutarate (HTK) solution. Murine syngeneic heterotopic heart transplantations (Balb/c-Balb/c) were carried out after 18 h of static cold storage. Cardiac grafts were either flushed and stored with Unisol-based solutions with high-(UHK) and low-potassium (ULK) ± glutathione, or HTK. Cardiac grafts were assessed for rebeating and functionality, histomorphologic alterations, and cytokine expression. Unisol-based solutions demonstrated a faster rebeating time (UHK 56 s, UHK + Glut 44 s, ULK 45 s, ULK + Glut 47 s) compared to HTK (119.5 s) along with a better contractility early after reperfusion and at the endpoint on POD 3. Ischemic injury led to a significantly increased leukocyte recruitment, with similar degrees of tissue damage and inflammatory infiltrate in all groups, yet the number of apoptotic cells tended to be lower in ULK compared to HTK. In UHK- and ULK-treated animals, a trend toward decreased expression of proinflammatory markers was seen when compared to HTK. Unisol-based solutions showed an improved preservative capacity compared with the gold standard HTK early after cardiac transplantation. Supplemented glutathione did not further improve tissue-protective properties. Full article
(This article belongs to the Special Issue Biobanking of Engineered and Natural Tissues)
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18 pages, 3822 KiB  
Article
Development of a Vitrification Preservation Process for Bioengineered Epithelial Constructs
by Lia H. Campbell and Kelvin G. M. Brockbank
Cells 2022, 11(7), 1115; https://doi.org/10.3390/cells11071115 - 25 Mar 2022
Cited by 5 | Viewed by 3133
Abstract
The demand for human bioengineered tissue constructs is growing in response to the worldwide movement away from the use of animals for testing of new chemicals, drug screening and household products. Presently, constructs are manufactured and delivered just in time, resulting in delays [...] Read more.
The demand for human bioengineered tissue constructs is growing in response to the worldwide movement away from the use of animals for testing of new chemicals, drug screening and household products. Presently, constructs are manufactured and delivered just in time, resulting in delays and high costs of manufacturing. Cryopreservation and banking would speed up delivery times and permit cost reduction due to larger scale manufacturing. Our objective in these studies was development of ice-free vitrification formulations and protocols using human bioengineered epithelial constructs that could be scaled up from individual constructs to 24-well plates. Initial experiments using single EpiDerm constructs in vials demonstrated viability >80% of untreated control, significantly higher than our best freezing strategy. Further studies focused on optimization and evaluation of ice-free vitrification strategies. Vitrification experiments with 55% (VS55) and 70% (VS70) cryoprotectant (CPA) formulations produced constructs with good viability shortly after rewarming, but viability decreased in the next days, post-rewarming in vitro. Protocol changes contributed to improved outcomes over time in vitro. We then transitioned from using glass vials with 1 construct to deep-well plates holding up to 24 individual constructs. Construct viability was maintained at >80% post-warming viability and >70% viability on days 1–3 in vitro. Similar viability was demonstrated for other related tissue constructs. Furthermore, we demonstrated maintenance of viability after 2–7 months of storage below −135 °C. Full article
(This article belongs to the Special Issue Biobanking of Engineered and Natural Tissues)
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17 pages, 2342 KiB  
Article
Assessment of the Impact of Post-Thaw Stress Pathway Modulation on Cell Recovery following Cryopreservation in a Hematopoietic Progenitor Cell Model
by John M. Baust, Kristi K. Snyder, Robert G. Van Buskirk and John G. Baust
Cells 2022, 11(2), 278; https://doi.org/10.3390/cells11020278 - 14 Jan 2022
Cited by 15 | Viewed by 3787
Abstract
The development and use of complex cell-based products in clinical and discovery science continues to grow at an unprecedented pace. To this end, cryopreservation plays a critical role, serving as an enabling process, providing on-demand access to biological material, facilitating large scale production, [...] Read more.
The development and use of complex cell-based products in clinical and discovery science continues to grow at an unprecedented pace. To this end, cryopreservation plays a critical role, serving as an enabling process, providing on-demand access to biological material, facilitating large scale production, storage, and distribution of living materials. Despite serving a critical role and substantial improvements over the last several decades, cryopreservation often remains a bottleneck impacting numerous areas including cell therapy, tissue engineering, and tissue banking. Studies have illustrated the impact and benefit of controlling cryopreservation-induced delayed-onset cell death (CIDOCD) through various “front end” strategies, such as specialized media, new cryoprotective agents, and molecular control during cryopreservation. While proving highly successful, a substantial level of cell death and loss of cell function remains associated with cryopreservation. Recently, we focused on developing technologies (RevitalICE™) designed to reduce the impact of CIDOCD through buffering the cell stress response during the post-thaw recovery phase in an effort to improve the recovery of previously cryopreserved samples. In this study, we investigated the impact of modulating apoptotic caspase activation, oxidative stress, unfolded protein response, and free radical damage in the initial 24 h post-thaw on overall cell survival. Human hematopoietic progenitor cells in vitro cryopreserved in both traditional extracellular-type and intracellular-type cryopreservation freeze media were utilized as a model cell system to assess impact on survival. Our findings demonstrated that through the modulation of several of these pathways, improvements in cell recovery were obtained, regardless of the freeze media and dimethyl sulfoxide concentration utilized. Specifically, through the use of oxidative stress inhibitors, an average increase of 20% in overall viability was observed. Furthermore, the results demonstrated that by using the post-thaw recovery reagent on samples cryopreserved in intracellular-type media (Unisol™), improvements in overall cell survival approaching 80% of non-frozen controls were attained. While improvements in overall survival were obtained, an assessment on the impact of specific cell subpopulations and functionality remains to be completed. While work remains, these results represent an important step forward in the development of improved cryopreservation processes for use in discovery science, and commercial and clinical settings. Full article
(This article belongs to the Special Issue Biobanking of Engineered and Natural Tissues)
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Review

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9 pages, 759 KiB  
Review
Cryopreservation by Directional Freezing and Vitrification Focusing on Large Tissues and Organs
by Amir Arav
Cells 2022, 11(7), 1072; https://doi.org/10.3390/cells11071072 - 22 Mar 2022
Cited by 15 | Viewed by 4749
Abstract
The cryopreservation of cells has been in routine use for decades. However, despite the extensive research in the field, cryopreservation of large tissues and organs is still experimental. The present review highlights the major studies of directional freezing and vitrification of large tissues [...] Read more.
The cryopreservation of cells has been in routine use for decades. However, despite the extensive research in the field, cryopreservation of large tissues and organs is still experimental. The present review highlights the major studies of directional freezing and vitrification of large tissues and whole organs and describes the different parameters that impact the success rate of large tissue and organ cryopreservation. Key factors, such as mass and heat transfer, cryoprotectant toxicity, nucleation, crystal growth, and chilling injury, which all have a significant influence on whole-organ cryopreservation outcomes, are reviewed. In addition, an overview of the principles of directional freezing and vitrification is given and the manners in which cryopreservation impacts large tissues and organs are described in detail. Full article
(This article belongs to the Special Issue Biobanking of Engineered and Natural Tissues)
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