Perfusion decellularization of whole organs
Abstract
The native extracellular matrix (ECM) outlines the architecture of organs and tissues. It provides a unique niche of composition and form, which serves as a foundational scaffold that supports organ-specific cell types and enables normal organ function. Here we describe a standard process for pressure-controlled perfusion decellularization of whole organs for generating acellular 3D scaffolds with preserved ECM protein content, architecture and perfusable vascular conduits. By applying antegrade perfusion of detergents and subsequent washes to arterial vasculature at low physiological pressures, successful decellularization of complex organs (i.e., hearts, lungs and kidneys) can be performed. By using appropriate modifications, pressure-controlled perfusion decellularization can be achieved in small-animal experimental models (rat organs, 4-5 d) and scaled to clinically relevant models (porcine and human organs, 12-14 d). Combining the unique structural and biochemical properties of native acellular scaffolds with subsequent recellularization techniques offers a novel platform for organ engineering and regeneration, for experimentation ex vivo and potential clinical application in vivo.
Integrity signals
Registry
Cites
Statistics
Claims the paper makes
Claims other papers make about this one
Each is checked against this paper’s own text: does it say what the citing paper says it does?
Some organoids have been grown in decellularized ECM from human or animal donors.
“In order to accurately recapitulate the composition, structure, and vascularization of native ECM in organ development, some organoids have been grown in decellularized ECM from human or animal donors.”
From Towards organoid culture without Matrigel (2021) · cites this paper as [70]
Decellularization approaches have been demonstrated for human kidney, murine kidney, murine heart, human and porcine lung, and porcine testicular tissues.
“Decellularization approaches have been demonstrated for human kidney67, murine kidney67,68, murine heart69,70, human and porcine lung71, and porcine testicular72 tissues, with each type posing unique challenges.”
From Towards organoid culture without Matrigel (2021) · cites this paper as [70]
Cites
An overview of tissue and whole organ decellularization processes
Crapo, P. M., Gilbert, T. W., & Badylak, S. F. (2011) · Biomaterials · doi:10.1016/j.biomaterials.2011.01.057 · Cited by 3,591 · see citing works
Perfusion-decellularized matrix: using nature's platform to engineer a bioartificial heart
Ott, H. C., Matthiesen, T. S., Goh, S.-K., Black, L. D., Kren, S. M., Netoff, T. I., & Taylor, D. A. (2008) · Nature Medicine · doi:10.1038/nm1684 · Cited by 2,650 · see citing works
Self-citationDecellularization of tissues and organs
Gilbert, T. W., Sellaro, T. L., & Badylak, S. F. (2006) · Biomaterials · doi:10.1016/j.biomaterials.2006.02.014 · Cited by 2,222 · see citing works
Yarbrough, W. M., Robbins, R. C., & Mallidi, H. R. (2016) · CRC Press eBooks · doi:10.3109/9781439802571-25 · Cited by 2,158 · see citing works
Extracellular matrix as a biological scaffold material: Structure and function
BADYLAK, S., FREYTES, D., & GILBERT, T. (2009) · Acta Biomaterialia · doi:10.1016/j.actbio.2008.09.013 · Cited by 1,671 · see citing works
Uygun, B. E., Soto-Gutierrez, A., Yagi, H., Izamis, M.-L., Guzzardi, M. A., Shulman, C., Milwid, J., Kobayashi, N., Tilles, A., Berthiaume, F., et al. (2010) · Nature Medicine · doi:10.1038/nm.2170 · Cited by 1,370 · see citing works
Reprint of: Extracellular matrix as a biological scaffold material: Structure and function
Badylak, S. F., Freytes, D. O., & Gilbert, T. W. (2015) · Acta Biomaterialia · doi:10.1016/j.actbio.2015.07.016 · Cited by 1,361 · see citing works
Tissue-Engineered Lungs for in Vivo Implantation
Petersen, T. H., Calle, E. A., Zhao, L., Lee, E. J., Gui, L., Raredon, M. S. B., Gavrilov, K., Yi, T., Zhuang, Z. W., Breuer, C. K., et al. (2010) · Science · doi:10.1126/science.1189345 · Cited by 1,177 · see citing works
Nelson, C. M., & Bissell, M. J. (2006) · Annual Review of Cell and Developmental Biology · doi:10.1146/annurev.cellbio.22.010305.104315 · Cited by 1,121 · see citing works
Regeneration and orthotopic transplantation of a bioartificial lung
Ott, H. C., Clippinger, B., Conrad, C., Schuetz, C., Pomerantseva, I., Ikonomou, L., Kotton, D., & Vacanti, J. P. (2010) · Nature Medicine · doi:10.1038/nm.2193 · Cited by 1,090 · see citing works
Self-citationBadylak, S. F., Taylor, D., & Uygun, K. (2011) · Annual Review of Biomedical Engineering · doi:10.1146/annurev-bioeng-071910-124743 · Cited by 1,041 · see citing works
Matricellular proteins: extracellular modulators of cell function
BORNSTEIN, P. K., & Sage, E. H. (2002) · Current Opinion in Cell Biology · doi:10.1016/s0955-0674(02)00361-7 · Cited by 966 · see citing works
Regeneration and experimental orthotopic transplantation of a bioengineered kidney
Song, J. J., Guyette, J. P., Gilpin, S. E., Gonzalez, G., Vacanti, J. P., & Ott, H. C. (2013) · Nature Medicine · doi:10.1038/nm.3154 · Cited by 793 · see citing works
Self-citationRieder, E., Kasimir, M.-T., Silberhumer, G., Seebacher, G., Wolner, E., Simon, P., & Weigel, G. (2004) · The Journal of Thoracic and Cardiovascular Surgery · doi:10.1016/j.jtcvs.2003.06.017 · Cited by 430 · see citing works
Mostow, E. N., Haraway, G. D., Dalsing, M. C., Hodde, J. P., & King, D. W. (2005) · Journal of Vascular Surgery · doi:10.1016/j.jvs.2005.01.042 · Cited by 387 · see citing works
Cortiella, J., Niles, J. A., Cantu, A., Brettler, A. C., Pham, A. N., Vargas, G., Winston, S., Wang, J. P., Walls, S., & Nichols, J. E. (2010) · Tissue Engineering Part A · doi:10.1089/ten.tea.2009.0730 · Cited by 381 · see citing works
Lu, T., Lin, B., Kim, J. K., Sullivan, M. L. G., Tobita, K., Salama, G., & Yang, L. (2013) · Nature Communications · doi:10.1038/ncomms3307 · Cited by 370 · see citing works
Decellularized tissue-engineered blood vessel as an arterial conduit
Quint, C., Kondo, Y., Manson, R. J., Lawson, J. H., Dardik, A., & Niklason, L. E. (2011) · Proceedings of the National Academy of Sciences · doi:10.1073/pnas.1019506108 · Cited by 368 · see citing works
Sullivan, D. C., Mirmalek-Sani, S.-H., Deegan, D. B., Baptista, P. M., Aboushwareb, T., Atala, A., & Yoo, J. J. (2012) · Biomaterials · doi:10.1016/j.biomaterials.2012.07.023 · Cited by 363 · see citing works
Price, A., England, K. A., Matson, A. M., Blazar, B. R., & Panoskaltsis‐Mortari, A. (2010) · Tissue Engineering Part A · doi:10.1089/ten.tea.2009.0659 · Cited by 344 · see citing works
Preparation of Cardiac Extracellular Matrix from an Intact Porcine Heart
Wainwright, J. M., Czajka, C. A., Patel, U. B., Freytes, D. O., Tobita, K., Gilbert, T. W., & Badylak, S. F. (2009) · Tissue Engineering Part C Methods · doi:10.1089/ten.tec.2009.0392 · Cited by 327 · see citing works
Decellularized matrices for tissue engineering
Hoshiba, T., Lu, H., Kawazoe, N., & Chen, G. (2010) · Expert Opinion on Biological Therapy · doi:10.1517/14712598.2010.534079 · Cited by 295 · see citing works
FLUID MECHANICS OF ARTIFICIAL HEART VALVES
Dasi, L. P., Simon, H., Sucosky, P., et al. (2009) · Clinical and Experimental Pharmacology and Physiology · doi:10.1111/j.1440-1681.2008.05099.x · Cited by 288 · see citing works
Goh, S.-K., Bertera, S., Olsen, P., Candiello, J., Halfter, W. M., Uechi, G. T., Balasubramani, M., Johnson, S. A., Sicari, B. M., Kollar, E. W., et al. (2013) · Biomaterials · doi:10.1016/j.biomaterials.2013.05.066 · Cited by 272 · see citing works
Cebotari, S., Tudorache, I., Jaekel, T., Hilfiker, A., Dorfman, S. E., Ternes, W., Haverich, A., & Lichtenberg, A. (2010) · Artificial Organs · doi:10.1111/j.1525-1594.2009.00796.x · Cited by 272 · see citing works
Perfusion decellularization of human and porcine lungs: Bringing the matrix to clinical scale
Gilpin, S. E., Guyette, J. P., Gonzalez, G., Ren, X., Asara, J. M., Mathisen, D. J., Vacanti, J. P., & Ott, H. C. (2014) · The Journal of Heart and Lung Transplantation · doi:10.1016/j.healun.2013.10.030 · Cited by 263 · see citing works
Self-citationDecellularization of Human and Porcine Lung Tissues for Pulmonary Tissue Engineering
O’Neill, J. D., Anfang, R. R., Anandappa, A. J., Costa, J. A., Javidfar, J., Wobma, H., Singh, G., Freytes, D. O., Bacchetta, M. D., Sonett, J. R., et al. (2013) · The Annals of Thoracic Surgery · doi:10.1016/j.athoracsur.2013.04.022 · Cited by 258 · see citing works
Orlando, G., Farney, A. C., Iskandar, S. S., Mirmalek-Sani, S.-H., Sullivan, D. C., Moran, E., AbouShwareb, T., De Coppi, P., Wood, K. J., Stratta, R. J., et al. (2012) · Annals of Surgery · doi:10.1097/sla.0b013e31825a02ab · Cited by 229 · see citing works
Extracellular Matrix Biomaterials for Soft Tissue Repair
Cornwell, K. G., Landsman, A. S., & James, K. S. (2009) · Clinics in Podiatric Medicine and Surgery · doi:10.1016/j.cpm.2009.08.001 · Cited by 215 · see citing works
Yang, B., Zhang, Y., Zhou, L., Sun, Z., Zheng, J., Chen, Y., & Dai, Y. (2010) · Tissue Engineering Part C: Methods · doi:10.1089/ten.tec.2009.0311 · Cited by 206 · see citing works
Wallis, J. M., Borg, Z. D., Daly, A. B., Deng, B., Ballif, B. A., Allen, G. B., Jaworski, D. M., & Weiss, D. J. (2011) · Tissue Engineering Part C Methods · doi:10.1089/ten.tec.2011.0567 · Cited by 187 · see citing works
Enhanced In Vivo Function of Bioartificial Lungs in Rats
Song, J. J., Kim, S. S., Liu, Z., Madsen, J. C., Mathisen, D. J., Vacanti, J. P., & Ott, H. C. (2011) · The Annals of Thoracic Surgery · doi:10.1016/j.athoracsur.2011.05.018 · Cited by 182 · see citing works
Self-citationActivation of Human Matrix Metalloproteinases by Various Bacterial Proteinases
Okamoto, T., Akaike, T., Suga, M., Tanase, S., Horie, H., Miyajima, S., Ando, M., Ichinose, Y., & Maeda, H. (1997) · Journal of Biological Chemistry · doi:10.1074/jbc.272.9.6059 · Cited by 177 · see citing works
Production and Assessment of Decellularized Pig and Human Lung Scaffolds
Nichols, J. E., Niles, J. A., Riddle, M., Vargas, G., Shilagard, T., Ma, L., Edward, K., Francesca, S. L., Sakamoto, J. H., Vega, S. P., et al. (2013) · Tissue Engineering Part A · doi:10.1089/ten.tea.2012.0250 · Cited by 166 · see citing works
Bonvillain, R. W., Danchuk, S. D., Sullivan, D. E., Betancourt, A. M., Semon, J. A., Eagle, M. E., Mayeux, J. P., Gregory, A. N., Wang, G., Townley, I. K., et al. (2012) · Tissue Engineering Part A · doi:10.1089/ten.tea.2011.0594 · Cited by 163 · see citing works
He, M., & Callanan, A. (2012) · Tissue Engineering Part B Reviews · doi:10.1089/ten.teb.2012.0340 · Cited by 160 · see citing works
Akhyari, P., Aubin, H., Gwanmesia, P., Barth, M., Hoffmann, S., Huelsmann, J., Preuß, K., & Lichtenberg, A. (2011) · Tissue Engineering Part C Methods · doi:10.1089/ten.tec.2011.0210 · Cited by 158 · see citing works
Effects of Freezing and Cryopreservation on the Mechanical Properties of Arteries
Venkatasubramanian, R. T., Grassl, E. D., Barocas, V. H., et al. (2006) · Annals of Biomedical Engineering · doi:10.1007/s10439-005-9044-x · Cited by 157 · see citing works
Freeze-Thaw Cycles Enhance Decellularization of Large Tendons
Burk, J., Erbe, I., Berner, D., Kacza, J., Kasper, C., Pfeiffer, B., Winter, K., & Brehm, W. (2013) · Tissue Engineering Part C Methods · doi:10.1089/ten.tec.2012.0760 · Cited by 151 · see citing works
Jungebluth, P., Go, T., Asnaghi, M. A., Bellini, S., Martorell, J., Calore, C., Urbani, L., Ostertag, H., Mantero, S., Conconi, M. T., et al. (2009) · Journal of Thoracic and Cardiovascular Surgery · doi:10.1016/j.jtcvs.2008.09.085 · Cited by 128 · see citing works
Ren, H., Shi, X., Tao, L., Xiao, J., Han, B., Zhang, Y., Yuan, X., & Ding, Y. (2013) · Liver International · doi:10.1111/liv.12088 · Cited by 127 · see citing works
Perspectives on whole-organ assembly: moving toward transplantation on demand
Soto–Gutiérrez, A., Wertheim, J. A., Ott, H. C., & Gilbert, T. W. (2012) · Journal of Clinical Investigation · doi:10.1172/jci61974 · Cited by 115 · see citing works
Self-citationEffects of freezing/thawing on the mechanical properties of decellularized lungs
Nonaka, P. N., Campillo, N., Uriarte, J. J., Garreta, E., Melo, E., de Oliveira, L. V. F., Navajas, D., & Farré, R. (2013) · Journal of Biomedical Materials Research Part A · doi:10.1002/jbm.a.34708 · Cited by 110 · see citing works
Yang, M., Chen, C., Wang, X., Zhu, Y., & Gu, Y. (2009) · Journal of Biomedical Materials Research Part B Applied Biomaterials · doi:10.1002/jbm.b.31409 · Cited by 108 · see citing works
Sokocevic, D., Bonenfant, N. R., Wagner, D. E., Borg, Z. D., Lathrop, M. J., Lam, Y. W., Deng, B., DeSarno, M., Ashikaga, T., Loi, R., et al. (2013) · Biomaterials · doi:10.1016/j.biomaterials.2013.01.028 · Cited by 99 · see citing works
Jensen, T., Roszell, B. R., Zang, F., Girard, E. D., Matson, A. P., Thrall, R. S., Jaworski, D. M., Hatton, C., Weiss, D. J., & Finck, C. M. (2012) · Tissue Engineering Part C Methods · doi:10.1089/ten.tec.2011.0584 · Cited by 93 · see citing works
Lähteenmäki, K., Virkola, R., Pouttu, R., Kuusela, P., Kukkonen, M., & Korhonen, T. K. (1995) · Infection and Immunity · doi:10.1128/iai.63.9.3659-3664.1995 · Cited by 87 · see citing works
Rusconi, F., Valton, É., Nguyen, R., & Dufourc, É. J. (2001) · Analytical Biochemistry · doi:10.1006/abio.2001.5164 · Cited by 84 · see citing works
Human Lung Cancer Cells Grown on Acellular Rat Lung Matrix Create Perfusable Tumor Nodules
Mishra, D. K., Thrall, M. J., Baird, B. N., Ott, H. C., Blackmon, S. H., Kurie, J. M., & Kim, M. P. (2012) · The Annals of Thoracic Surgery · doi:10.1016/j.athoracsur.2012.01.011 · Cited by 77 · see citing works
Self-citationVenkatasubramanian, R. T., Wolkers, W. F., Shenoi, M. M., et al. (2010) · Annals of Biomedical Engineering · doi:10.1007/s10439-010-9921-9 · Cited by 72 · see citing works
Boyd, W. D., Johnson, W. E. B., Sultan, P. K., Deering, T. F., & Matheny, R. G. (2010) · The Heart Surgery Forum · doi:10.1532/hsf98.20091184 · Cited by 47 · see citing works
Conrad, C., Schuetz, C., Clippinger, B. B., et al. (2010) · Journal of the American College of Surgeons · doi:10.1016/j.jamcollsurg.2010.06.161 · Cited by 26 · see citing works
Lee, L. T., Deas, J. E., & Howe, C. (1978) · Journal of Immunological Methods · doi:10.1016/0022-1759(78)90009-1 · Cited by 23 · see citing works
Dewaxed ECM: A simple method for analyzing cell behaviour on decellularized extracellular matrices
Ofenbauer, A., Sebinger, D., Prewitz, M. C., Gruber, P., & Werner, C. (2012) · Journal of Tissue Engineering and Regenerative Medicine · doi:10.1002/term.1658 · Cited by 15 · see citing works
Terada, H., Hideaki, S. E. K. I., Yamamoto, K., et al. (1985) · Analytical Biochemistry · doi:10.1016/0003-2697(85)90605-0 · Cited by 8 · see citing works
Flagged first, then by citations
Cited by
Sorted by citations
Decellularized human liver as a natural 3D-scaffold for liver bioengineering and transplantation
Mazza, G., Rombouts, K., Rennie Hall, A., Urbani, L., Vinh Luong, T., Al-Akkad, W., Longato, L., Brown, D., Maghsoudlou, P., Dhillon, A. P., et al. (2015) · Scientific Reports · doi:10.1038/srep13079 · Cited by 415 · see citing works
Towards organoid culture without Matrigel
Kozlowski, M. T., Crook, C. J., & Ku, H. T. (2021) · Communications Biology · doi:10.1038/s42003-021-02910-8 · Cited by 409 · see citing works
Bioengineering human myocardium on native extracellular matrix
Guyette, J. P., Charest, J. M., Mills, R. W., Jank, B. J., Moser, P. T., Gilpin, S. E., Gershlak, J. R., Okamoto, T., Gonzalez, G., Milan, D. J., et al. (2016) · Circulation Research · doi:10.1161/circresaha.115.306874 · Cited by 343 · see citing works
Self-citationRana, D., Zreiqat, H., Benkirane-Jessel, N., Ramakrishna, S., & Ramalingam, M. (2017) · Journal of Tissue Engineering and Regenerative Medicine · doi:10.1002/term.2061 · Cited by 243 · see citing works
Bioengineering Approaches for the Advanced Organoid Research
Yi, S. A., Zhang, Y., Rathnam, C., Pongkulapa, T., & Lee, K. (2021) · Advanced Materials · doi:10.1002/adma.202007949 · Cited by 218 · see citing works
Fibrous scaffolds for building hearts and heart parts
Capulli, A. K., MacQueen, L. A., Sheehy, S. P., & Parker, K. K. (2016) · Advanced Drug Delivery Reviews · doi:10.1016/j.addr.2015.11.020 · Cited by 130 · see citing works
Grattoni, A., Korbutt, G., Tomei, A. A., García, A. J., Pepper, A. R., Stabler, C., Brehm, M., Papas, K., Citro, A., Shirwan, H., et al. (2025) · Nature Reviews Endocrinology · doi:10.1038/s41574-024-01029-0 · Cited by 92 · see citing works
Messner, F., Guo, Y., Etra, J. W., & Brandacher, G. (2019) · Transplant International · doi:10.1111/tri.13432 · Cited by 29 · see citing works
Immune Response to Extracellular Matrix Bioscaffolds: A Comprehensive Review
Romero, D. J., Hussey, G., & Capella-Monsonís, H. (2025) · Biologics · doi:10.3390/biologics5030028 · Cited by 7 · see citing works
Strategies for decellularization, re-cellularization and crosslinking in liver bioengineering
Wang, J., & Jin, X. (2024) · The International Journal of Artificial Organs · doi:10.1177/03913988231218566 · Cited by 3 · see citing works
- Authors
- Guyette, J. P., Gilpin, S. E., Charest, J. M., Tapias, L. F., Ren, X., & Ott, H. C.
- Affiliations
- Harvard University (US)
- Massachusetts General Hospital (US)
- Corresponding author
- Ott, Harald C
- Venue
- Nature Protocols
- Published
- 2014-05-29
- Volume
- 9
- Issue
- 6
- Pages
- 1451-1468
- ISSN
- 1750-2799 (ISSN-L)
- 1754-2189
- OpenAlex
- W2068102665
- Cited by
- 275
- References
- 55
- Citations by year
- 2026: 11 · 2025: 15 · 2024: 23 · 2023: 29 · 2022: 18 · 2021: 29 · 2020: 17 · 2019: 26 · 2018: 27 · 2017: 30 · 2016: 17 · 2015: 30 · 2014: 3
- FWCI
- 26.43
- Type
- journal article
- Language
- en
- Open access
- closed
- Locations
- Nature Protocols (publishedVersion)
- PubMed (publishedVersion)
- Funders
- National Heart, Lung, and Blood Institute (R21HL108663, R01HL108678)
- Topics
- Health Sciences › Medicine › Surgery › Tissue Engineering and Regenerative Medicine
- Physical Sciences › Engineering › Biomedical Engineering › 3D Printing in Biomedical Research
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Medicine › Hydrogels: synthesis, properties, applications
- Keywords
- perfusion decellularization, extracellular matrix, vascular conduits, detergents, antegrade perfusion, heart, lung, kidney, recellularization, organ engineering, tissue regeneration
- MeSH terms
- Animals, Detergents, Extracellular Matrix, Humans, Perfusion, Pressure, Swine, Viscera, Extracellular Matrix Proteins, Tissue Engineering, Rats, Tissue Scaffolds
About these checks
Registry, retraction, Cites and statistics results are computed, not judged, with one exception: a printed reference that no index lists is read once by a model before it counts as a citation error. Claims are extracted by a model and kept only when their quote appears verbatim in the text; each cited source is then checked, with a model’s help, for whether it says what the claim relies on. Those verdicts can be wrong — read the claim’s quote and the cited source before relying on one.
Statistics and arithmetic are recomputed, not judged.
Confidence bands are shown only for fields calibrated on our annotated test set. None is calibrated yet, so no band is shown.
What counts as a claim ›How we measure confidence ›How we check this ›
Are you an author, or do you see a problem with this page? Request a review
