Intravital microscopy datasets examining key nephron segments of transplanted decellularized kidneys

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Abstract

This study contains intravital microscopy (IVM) data examining the microarchitecture of acellular kidney scaffolds. Acellular scaffolds are cell-free collagen-based matrices derived from native organs that can be used as templates for regenerative medicine applications. This data set contains in vivo assays that evaluate the effectiveness of decellularization and how these acellular nephron compartments perform in the post-transplantation environment. Qualitative and quantitative assessments of scaffold DNA concentrations, tissue fluorescence signals, and structural and functional integrities of decellularized tubular and peritubular capillary segments were acquired and compared to the native (non-transplanted) organ. Cohorts of 2–3-month-old male Sprague Dawley rats were used: non-transplanted (n = 4), transplanted day 0 (n = 4), transplanted day 1 (n = 4), transplanted day 2 (n = 4), and transplanted day 7 (n = 4). Micrographs and supporting measurements are provided to illustrate IVM processes used to perform this study and are publicly available in a data repository to assist scientific reproducibility and extend the use of this powerful imaging application to analyze other scaffold systems.

Measurements(s)	DNA quantification • tissue fluorescence • microvascular leakage • tubular and peritubular capillary integrity
Technology Type(s)	intravital microscopy • multiphoton microscopy • UV-visible spectroscopy
Sample Characterization(s)	rats • native and decellularized kidneys

Abstract

Measurement(s)	DNA • tissue fluorescence
Technology Type(s)	ultraviolet-visible spectrophotometry • intravital microscopy
Sample Characteristic - Organism	mouse/rat NG108-15
Sample Characteristic - Environment	laboratory environment
Sample Characteristic - Location	contiguous United States of America • Emirate (United Arab Emirates)

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Decellularization Strategies for Regenerative Medicine: From Processing Techniques to Applications

Anna Gilpin, Yong Yang (2017)

As the gap between donors and patients in need of an organ transplant continues to widen, research in regenerative medicine seeks to provide alternative strategies for treatment. One of the most promising techniques for tissue and organ regeneration is decellularization, in which the extracellular matrix (ECM) is isolated from its native cells and genetic material in order to produce a natural scaffold. The ECM, which ideally retains its inherent structural, biochemical, and biomechanical cues, can then be recellularized to produce a functional tissue or organ. While decellularization can be accomplished using chemical and enzymatic, physical, or combinative methods, each strategy has both benefits and drawbacks. The focus of this review is to compare the advantages and disadvantages of these methods in terms of their ability to retain desired ECM characteristics for particular tissues and organs. Additionally, a few applications of constructs engineered using decellularized cell sheets, tissues, and whole organs are discussed.

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Preparation and characterization of decellularized cornea using high-hydrostatic pressurization for corneal tissue engineering.

Yoshihide Hashimoto, Seiichi Funamoto, Shuji Sasaki … (2010)

To prepare acellular corneal scaffold, we used high-hydrostatic pressurization (HHP) to decellularize porcine cornea. The HHP method disrupts cells by hydrostatic pressurization, and then the disrupted cells' components are removed by washing with a cell culture medium. Porcine corneas were hydrostatically pressed at 980 MPa at 10 or 30 degrees C for 10 min to make them opaque. There was no change in the thickness of the corneas immediately after the pressurization, but they swelled during the washing process. The cornea swelling caused by HHP was suppressed when medium containing 3.5% w/v dextran was used. For H-E staining of the cornea decellularized with the HHP method, the complete removal of corneal cells was confirmed. Furthermore, when the corneas were immersed in glycerol for 1 hour, their optical properties were restored to those of native corneas. In an animal study, when acellular porcine corneas were implanted into rabbit cornea, no immune reaction occurred and the turbid corneas became clear. The decellularized corneas obtained through HHP could be useful as a corneal scaffold for tissue regeneration. Copyright 2010 Elsevier Ltd. All rights reserved.

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The impact of detergents on the tissue decellularization process: A ToF-SIMS study.

Lisa J White, Adam J Taylor, Denver M Faulk … (2017)

Biologic scaffolds are derived from mammalian tissues, which must be decellularized to remove cellular antigens that would otherwise incite an adverse immune response. Although widely used clinically, the optimum balance between cell removal and the disruption of matrix architecture and surface ligand landscape remains a considerable challenge. Here we describe the use of time of flight secondary ion mass spectroscopy (ToF-SIMS) to provide sensitive, molecular specific, localized analysis of detergent decellularized biologic scaffolds. We detected residual detergent fragments, specifically from Triton X-100, sodium deoxycholate and sodium dodecyl sulphate (SDS) in decellularized scaffolds; increased SDS concentrations from 0.1% to 1.0% increased both the intensity of SDS fragments and adverse cell outcomes. We also identified cellular remnants, by detecting phosphate and phosphocholine ions in PAA and CHAPS decellularized scaffolds. The present study demonstrates ToF-SIMS is not only a powerful tool for characterization of biologic scaffold surface molecular functionality, but also enables sensitive assessment of decellularization efficacy.

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Author and article information

Contributors

Peter R. Corridon:

ORCID: http://orcid.org/0000-0002-6796-4301

peter.corridon@ku.ac.ae

Journal

Journal ID (nlm-ta): Sci Data

Journal ID (iso-abbrev): Sci Data

Title: Scientific Data

Publisher: Nature Publishing Group UK (London )

ISSN (Electronic): 2052-4463

Publication date (Electronic): 10 September 2022

Publication date PMC-release: 10 September 2022

Publication date Collection: 2022

Volume: 9

Electronic Location Identifier: 561

Affiliations

[1 ]GRID grid.440568.b, ISNI 0000 0004 1762 9729, Department of Immunology and Physiology, College of Medicine and Health Sciences, , Khalifa University of Science and Technology, ; PO Box 127788, Abu Dhabi, UAE

[2 ]GRID grid.412860.9, ISNI 0000 0004 0459 1231, Wake Forest Institute for Regenerative Medicine, , Medical Center Boulevard, ; Winston-Salem, NC 27157-1083 USA

[3 ]GRID grid.440568.b, ISNI 0000 0004 1762 9729, Biomedical Engineering, Healthcare Engineering Innovation Center, , Khalifa University of Science and Technology, ; PO Box 127788, Abu Dhabi, UAE

[4 ]GRID grid.440568.b, ISNI 0000 0004 1762 9729, Center for Biotechnology, , Khalifa University of Science and Technology, ; PO Box 127788, Abu Dhabi, UAE

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Peter R. Corridon http://orcid.org/0000-0002-6796-4301

Article

Publisher ID: 1685

DOI: 10.1038/s41597-022-01685-9

PMC ID: 9464233

SO-VID: be3020e6-bbe3-4cdc-814c-f70746eff5b1

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Open Access This article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made. The images or other third party material in this article are included in the article’s Creative Commons license, unless indicated otherwise in a credit line to the material. If material is not included in the article’s Creative Commons license and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this license, visit http://creativecommons.org/licenses/by/4.0/.

History

Date received : 19 April 2022

Date accepted : 7 September 2022

Funding

Funded by: FundRef https://doi.org/10.13039/100000057, U.S. Department of Health & Human Services | NIH | National Institute of General Medical Sciences (NIGMS);

Award ID: NIH/NIGMS K12-GM102773

Award Recipient : Peter R. Corridon

Funded by: FundRef https://doi.org/10.13039/501100004070, Khalifa University of Science, Technology and Research (Khalifa University);

Award ID: FSU-2020-25

Award ID: RC2-2018-022 (HEIC)

Award Recipient : Peter R. Corridon

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Keywords: end-stage renal disease,biological fluorescence,multiphoton microscopy

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Keywords: end-stage renal disease, biological fluorescence, multiphoton microscopy

Intravital microscopy datasets examining key nephron segments of transplanted decellularized kidneys

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Most cited references 37

Decellularization Strategies for Regenerative Medicine: From Processing Techniques to Applications

Preparation and characterization of decellularized cornea using high-hydrostatic pressurization for corneal tissue engineering.

The impact of detergents on the tissue decellularization process: A ToF-SIMS study.

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