Name: Author Spotlight: Optimizing Bovine Lung Decellularization for Organotypic Hydrogels
Uploaded: 2023-12-08T13:10:00
Description: Watch this Scientific Journal Video about Development and Characterization of Decellularized Lung Extracellular Matrix Hydrogels at JoVE.com

This work was funded by the Scientific and Technological Research Council of Turkey (T&#220;B&#304;TAK) (Grant No. 118C238). The entire responsibility of the publication/paper belongs to the owner of the publication. The financial support received from TU&#776;BI&#775;TAK does not mean that the content of the publication is approved in a scientific sense by TU&#776;BI&#775;TAK. The authors gratefully acknowledge the use of services and facilities of Koc&#807; University Research Center for Translational Medicine (KUTTAM). Figure 1 and Figure 2a were created using Biorender.com.

Fresh native lungs from young (1-2 years old) bovine donors were obtained from a local slaughterhouse and transported in a sealed plastic container on ice to the laboratory. Animal sacrifice is performed for general meat consumption (lungs discarded as waste) and is not related to or due to the study. We confirm that the slaughterhouse complies with the national laws and regulations of animal sacrifice. Furthermore, we confirm that we only used waste material and the research project did not have an effect on the number ...

Comparative Study of Decellularization Techniques for Bovine Lung ECM Hydrogels

<ol>
	<li>Zhu, X., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Ordered+micropattern+arrays+fabricated+by+lung-derived+dECM+hydrogels+for+chemotherapeutic+drug+screening.">Ordered micropattern arrays fabricated by lung-derived dECM hydrogels for chemotherapeutic drug screening.</a> Mater Today Bio. 15, 100274 (2022).</li><li>Ulldemolins, A., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Lung+extracellular+matrix+hydrogels-derived+vesicles+contribute+to+epithelial+lung+repair.">Lung extracellular matrix hydrogels-derived vesicles contribute to epithelial lung repair.</a> Polymers. 14 (22), 4907 (2022).</li><li>Biehl, A., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Towards+a+standardized+multi-tissue+decellularization+protocol+for+the+derivation+of+extracellular+matrix+materials.">Towards a standardized multi-tissue decellularization protocol for the derivation of extracellular matrix materials.</a> Biomater Sci. 11 (2), 641-654 (2023).</li><li>Pak, J., Lee, J. H., Park, K. S., Jeong, B. C., Lee, S. H. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Regeneration+of+cartilage+in+human+knee+osteoarthritis+with+autologous+adipose+tissue-derived+stem+cells+and+autologous+extracellular+matrix.">Regeneration of cartilage in human knee osteoarthritis with autologous adipose tissue-derived stem cells and autologous extracellular matrix.</a> Biores Open Access. 5 (1), 192-200 (2016).</li><li>Spang, M. T., Christman, K. L. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Extracellular+matrix+hydrogel+therapies.">Extracellular matrix hydrogel therapies.</a> In vivo applications and development.Acta Biomater. 68, 1-14 (2018).</li><li>Schwartz, A. D., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=A+biomaterial+screening+approach+reveals+microenvironmental+mechanisms+of+drug+resistance.">A biomaterial screening approach reveals microenvironmental mechanisms of drug resistance.</a> Integr Biol. 9 (12), 912-924 (2017).</li><li>Monteiro, M. V., Gaspar, V. M., Ferreira, L. P., Mano, J. F. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Hydrogel+3D+in+vitro+tumor+models+for+screening+cell+aggregation+mediated+drug+response.">Hydrogel 3D in vitro tumor models for screening cell aggregation mediated drug response.</a> Biomater Sci. 8 (7), 1855-1864 (2020).</li><li>Nakamura, R., Nakamura, F., Fukunaga, S. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Changes+in+the+composition+of+the+extracellular+matrix+accumulated+by+mesenchymal+stem+cells+during+in+vitro+expansion.">Changes in the composition of the extracellular matrix accumulated by mesenchymal stem cells during in vitro expansion.</a> Anim Sci J. 85 (6), 706-713 (2014).</li><li>Bual, R. P., Ijima, H. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Intact+extracellular+matrix+component+promotes+maintenance+of+liver-specific+functions+and+larger+aggregates+formation+of+primary+rat+hepatocytes.">Intact extracellular matrix component promotes maintenance of liver-specific functions and larger aggregates formation of primary rat hepatocytes.</a> Regen Ther. 11, 258-268 (2019).</li><li>Jhala, D., Vasita, R. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=A+review+on+extracellular+matrix+mimicking+strategies+for+an+artificial+stem+cell+niche.">A review on extracellular matrix mimicking strategies for an artificial stem cell niche.</a> Poly Rev. 55 (4), 561-595 (2015).</li><li>Jeong, W., Kim, M. K., Kang, H. W. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Effect+of+detergent+type+on+the+performance+of+liver+decellularized+extracellular+matrix-based+bio-inks.">Effect of detergent type on the performance of liver decellularized extracellular matrix-based bio-inks.</a> J Tissue Eng. 12, 2041731421997091 (2021).</li><li>Lee, S. J., Lee, J. H., Park, J., Kim, W. D., Park, S. A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Fabrication+of+3D+printing+scaffold+with+porcine+skin+decellularized+bio-ink+for+soft+tissue+engineering.">Fabrication of 3D printing scaffold with porcine skin decellularized bio-ink for soft tissue engineering.</a> Materials. 13 (16), 3522 (2020).</li><li>Won, J. Y., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=A+potential+dermal+substitute+using+decellularized+dermis+extracellular+matrix+derived+bio-ink.">A potential dermal substitute using decellularized dermis extracellular matrix derived bio-ink.</a> Artif Cells Nanomed Biotechnol. 47 (1), 644-649 (2019).</li><li>Lin, Z., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Bioactive+decellularized+extracellular+matrix+hydrogel+microspheres+fabricated+using+a+temperature-controlling+microfluidic+system.">Bioactive decellularized extracellular matrix hydrogel microspheres fabricated using a temperature-controlling microfluidic system.</a> ACS Biomater Sci Eng. 8 (4), 1644-1655 (2022).</li><li>Kaushik, N., Kim, S., Suh, Y., Lee, S. J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Proinvasive+extracellular+matrix+remodeling+for+tumor+progression.">Proinvasive extracellular matrix remodeling for tumor progression.</a> Arch Pharm Res. 42 (1), 40-47 (2019).</li><li>Lu, P., Weaver, V. M., Werb, Z. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=The+extracellular+matrix:+a+dynamic+niche+in+cancer+progression.">The extracellular matrix: a dynamic niche in cancer progression.</a> J Cell Biol. 196 (4), 395-406 (2012).</li><li>Poltavets, V., Kochetkova, M., Pitson, S. M., Samuel, M. S. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=The+role+of+the+extracellular+matrix+and+its+molecular+and+cellular+regulators+in+cancer+cell+plasticity.">The role of the extracellular matrix and its molecular and cellular regulators in cancer cell plasticity.</a> Front Oncol. 8, 431 (2018).</li><li>Takeda, M., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Development+of+a+drug+screening+system+using+three-dimensional+cardiac+tissues+containing+multiple+cell+types.">Development of a drug screening system using three-dimensional cardiac tissues containing multiple cell types.</a> Sci Rep. 11 (1), 5654 (2021).</li><li>Jensen, A. R. D., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Organ-specific,+fibroblast-derived+matrix+as+a+tool+for+studying+breast+cancer+metastasis.">Organ-specific, fibroblast-derived matrix as a tool for studying breast cancer metastasis.</a> Cancers (Basel). 13 (13), 3331 (2021).</li><li>Ku&#351;o&#287;lu, A., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Different+decellularization+methods+in+bovine+lung+tissue+reveals+distinct+biochemical+composition,+stiffness,+and+viscoelasticity+in+reconstituted+hydrogels.">Different decellularization methods in bovine lung tissue reveals distinct biochemical composition, stiffness, and viscoelasticity in reconstituted hydrogels.</a> ACS Appl Bio Mater. 6 (2), 793-805 (2023).</li><li>Fern&#225;ndez-P&#233;rez, J., Ahearne, M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=The+impact+of+decellularization+methods+on+extracellular+matrix+derived+hydrogels.">The impact of decellularization methods on extracellular matrix derived hydrogels.</a> Sci Rep. 9 (1), 14933 (2019).</li><li>Park, S., Kim, T. H., Kim, S. H., You, S., Jung, Y. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Three-dimensional+vascularized+lung+cancer-on-a-chip+with+lung+extracellular+matrix+hydrogels+for+in+vitro+screening.">Three-dimensional vascularized lung cancer-on-a-chip with lung extracellular matrix hydrogels for in vitro screening.</a> Cancers (Basel). 13 (16), 3930 (2021).</li><li>Uriarte, J. J., Uhl, F. E., Rolandsson Enes, S. E., Pouliot, R. A., Weiss, D. J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Lung+bioengineering:+advances+and+challenges+in+lung+decellularization+and+recellularization.">Lung bioengineering: advances and challenges in lung decellularization and recellularization.</a> Curr Opin Organ Transplant. 23 (6), 673-678 (2018).</li><li>Roth, S. P., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Automated+freeze-thaw+cycles+for+decellularization+of+tendon+tissue+-+a+pilot+study.">Automated freeze-thaw cycles for decellularization of tendon tissue - a pilot study.</a> BMC Biotechnol. 17 (1), 13 (2017).</li><li>Gilbert, T. W., Freund, J. M., Badylak, S. F. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Quantification+of+DNA+in+biologic+scaffold+materials.">Quantification of DNA in biologic scaffold materials.</a> J Surg Res. 152 (1), 135-139 (2009).</li><li>Crapo, P. M., Gilbert, T. W., Badylak, S. F. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=An+overview+of+tissue+and+whole+organ+decellularization+processes.">An overview of tissue and whole organ decellularization processes.</a> Biomaterials. 32 (12), 3233-3243 (2011).</li><li>Cox, T. R. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=The+matrix+in+cancer.">The matrix in cancer.</a> Nat Rev Cancer. 21 (4), 217-238 (2021).</li><li>Vining, K. H., Mooney, D. J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Mechanical+forces+direct+stem+cell+behaviour+in+development+and+regeneration.">Mechanical forces direct stem cell behaviour in development and regeneration.</a> Nat Rev Mol Cell Biol. 18 (12), 728-742 (2017).</li><li>Chaudhuri, O., Cooper-White, J., Janmey, P. A., Mooney, D. J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Effects+of+extracellular+matrix+viscoelasticity+on+cellular+behaviour.">Effects of extracellular matrix viscoelasticity on cellular behaviour.</a> Nature. 584 (7822), 535-546 (2020).</li><li>Chaudhuri, O., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Hydrogels+with+tunable+stress+relaxation+regulate+stem+cell+fate+and+activity.">Hydrogels with tunable stress relaxation regulate stem cell fate and activity.</a> Nat Mater. 15 (3), 326-334 (2016).</li></ol>

Organ-derived hydrogels have become promising models that recapitulate the native tissue ECM and mimic organotypic cellular function. Although decellularized lung ECM has often been used in tissue engineering, a thorough characterization of biomaterial composition and mechanical properties will benefit a better understanding of how cell-ECM interactions can be modulated for modeling biological processes during homeostasis or disease. Particularly, assessment and control of mechanical properties of reconstituted hydrogels...

Conventional monolayer culture conditions do not offer a faithful representation of native tissue microenvironments and lack the ability to provide a three-dimensional (3D) scaffold with instructive ligands that enable cell-matrix and cell-cell interactions1. Extracellular matrix (ECM) composition and mechanical properties are highly tissue-specific, time-dependent, and undergo alterations in pathological conditions. Therefore, there is a need for biomimetic 3D tissue models that allow tunability of such characteristics, modulation of cellular behavior, and achieving desired tissue functionality. Native ECM-derived biomaterials draw much attention in tissue engineering with the ability to directly use tissue-specific ECM1,2,3,4,5. ECM-based carriers have been used in many applications spanning tissue regeneration to disease model development. They are used as injectable or implantable biomaterial scaffolds4,5, in drug screening applications6,7, in the development of materials that induce cell growth8,9,10, as bio-inks11,12,13, in microfluidics14, and in cancer tissue models15,16,17,18,19.
Decellularization of tissues and organs is a popular approach for&#160;generating scaffolds that mimic tissue-specific ECM. The reconstitution of decellularized tissues and organs into hydrogels allows embedding of cells into biomimetic 3D tissue models20. Decellularization techniques mainly focus on eliminating cellular components while retaining the ECM composition. Physical methods such as freeze-thaw cycles or chemical processes such as Triton-X-100 treatment are commonly&#160;applied to decellularize tissues. Furthermore, DNase treatment is preferred for removing residual DNA to minimize the immunological responses upon cell embedding. It is critical to achieve maximal cell removal and minimal ECM impairment to optimize decellularization procedures21. Besides these aspects, the characterization of reconstituted scaffolds&#39; biochemical and mechanical properties, including viscoelasticity and stiffness, is crucial for&#160;improving engineered 3D tissue models derived from native matrices20.
Organ-specific ECM in lung tissue engineering allows mimicking the pulmonary microenvironment to model developmental, homeostatic, or pathological processes in vitro and testing therapeutics in a physio-mimetic setting20,22,23. Previous studies have demonstrated decellularization of&#160;lung tissue from several species, such as rats, porcine, and humans, but these methods have yet to be adapted to less frequently used species such as bovine. A better understanding of the parameters of the decellularization process and how they affect the resulting reconstituted ECM scaffolds regarding biochemical composition and mechanical properties will allow for better tuning of such aspects and pave the way for more reliable tissue models in health and disease. In this study, bovine lung decellularization with two distinct methods, freeze-thaw cycles and Triton-X-100 treatment, is explicitly described and followed by biochemical and mechanical analyses of decellularized lung ECM (dECM) hydrogels. The findings reveal that both methods yield effective decellularization and retention of ECM ligands. Notably, the choice of method significantly alters the resulting stiffness and viscoelasticity of reconstituted hydrogels. Hydrogels derived from the bovine dECM demonstrate notable biochemical analogies with the extracellular matrix of the human lung, and they exhibit reliable thermal gelation characteristics20. As previously described, both methods are suitable for the 3D culture of lung cancer cells, healthy bronchial epithelial cells, and patient-derived lung organoids20.

<table border="1" fo:keep-together.within-page="1" fo:keep-with-next.within-page="always">
	<tbody>
		<tr>
			<td>Absolute ethanol</td>
			<td>ISOLAB</td>
			<td>64-17-5</td>
			<td></td>
		</tr>
		<tr>
			<td>Acetic acid</td>
			<td>ISOLAB</td>
			<td>64-19-7</td>
			<td></td>
		</tr>
		<tr>
			<td>Alcian blue solution</td>
			<td>Sigma-Aldrich</td>
			<td>B8438</td>
			<td></td>
		</tr>
		<tr>
			<td>Deoxyribonuclease I from bovine pancreas</td>
			<td>Sigma-Aldrich</td>
			<td>DN25</td>
			<td></td>
		</tr>
		<tr>
			<td>Discovery HR-2 rheometer</td>
			<td>TA Instruments</td>
			<td></td>
			<td></td>
		</tr>
		<tr>
			<td>Entellan mounting medium</td>
			<td>Merck</td>
			<td>107960</td>
			<td></td>
		</tr>
		<tr>
			<td>Eosin solution</td>
			<td>Bright-slide</td>
			<td>2.BS01-105-1000</td>
			<td></td>
		</tr>
		<tr>
			<td>Formaldehyde</td>
			<td>Electron Microscopy Sciences</td>
			<td>50-980-485</td>
			<td></td>
		</tr>
		<tr>
			<td>Hydrochloric acid</td>
			<td>Merck</td>
			<td>100317</td>
			<td></td>
		</tr>
		<tr>
			<td>Iodine</td>
			<td>Sigma-Aldrich</td>
			<td>3002</td>
			<td></td>
		</tr>
		<tr>
			<td>Magnesium chloride</td>
			<td>Sigma-Aldrich</td>
			<td>7786-30-3</td>
			<td></td>
		</tr>
		<tr>
			<td>Mayer&#39;s haematoxylin staining solution</td>
			<td>Merck</td>
			<td>2.BS01-103-1000</td>
			<td></td>
		</tr>
		<tr>
			<td>O.C.T compound</td>
			<td>Tissue-Tek</td>
			<td>4583</td>
			<td></td>
		</tr>
		<tr>
			<td>Penicillin/Streptomycin</td>
			<td>Biowest</td>
			<td>L0018-100</td>
			<td></td>
		</tr>
		<tr>
			<td>Pepsin from porcine gastric mucosa</td>
			<td>Sigma-Aldrich</td>
			<td>P6887</td>
			<td></td>
		</tr>
		<tr>
			<td>Picric acid</td>
			<td>Polysciences</td>
			<td>88-89-1</td>
			<td></td>
		</tr>
		<tr>
			<td>Sirius Red</td>
			<td>Polysciences</td>
			<td>09400-25</td>
			<td></td>
		</tr>
		<tr>
			<td>Sodium hydroxide</td>
			<td>Sigma-Aldrich</td>
			<td>S5881</td>
			<td></td>
		</tr>
		<tr>
			<td>Sucrose&#160;</td>
			<td>Sigma-Aldrich</td>
			<td>S0389</td>
			<td></td>
		</tr>
		<tr>
			<td>Triton-X-100</td>
			<td>Merck</td>
			<td>112298</td>
			<td></td>
		</tr>
	</tbody>
</table>

development and characterization of decellularized lung extracellular matrix hydrogels

The use of extracellular matrix (ECM)-derived hydrogels in tissue engineering has become increasingly popular, as they can mimic cells&#39; natural environment in vitro. However, maintaining the native biochemical content of the ECM, achieving mechanical stability, and comprehending the impact of the decellularization process on the mechanical properties of the ECM hydrogels are challenging. Here, a pipeline for decellularization of bovine lung tissue using two different protocols, downstream characterization of the effectiveness of decellularization, fabrication of reconstituted decellularized lung ECM hydrogels and assessment of their mechanical and cytocompatibility properties were described. Decellularization of the bovine lung was pursued using a physical (freeze-thaw cycles) or chemical (detergent-based) method. Hematoxylin and Eosin staining was performed to validate the decellularization and retention of major ECM components. For the evaluation of residual collagen and sulfated glycosaminoglycan (sGAG) content within the decellularized samples, Sirius red and Alcian blue staining techniques were employed, respectively. Mechanical properties of the decellularized lung ECM hydrogels were characterized by oscillatory rheology. The results suggest that decellularized bovine lung hydrogels can provide a reliable organotypic<span style="font-size: 11pt; font-family: Calibri, sans-serif; color: black; background-image: initial; background-position: initial; background-size: initial; background-repeat: initial; background-attachment: initial; background-origin: initial; background-clip: initial;">&#160;alternative to commercial ECM products by retaining most native ECM components. Furthermore, these findings reveal that the decellularization method of choice significantly affects gelation kinetics as well as the stiffness and viscoelastic properties of resulting hydrogels.

Author Spotlight: Optimizing Bovine Lung Decellularization for Organotypic Hydrogels

Development and Characterization of Decellularized Lung Extracellular Matrix Hydrogels

Decellularization 
Decellularization of bovine lung tissue to produce dECM hydrogels that would recapitulate the native lung microenvironment has been achieved by both physical (freeze-thaw) and chemical (Triton-X-100) methods. After dissection, tissue pieces were washed in dH2O-containing antibiotics to remove pathogens that can later affect the sterility of the dECM hydrogels. A total of five cycles alternating between liquid nitrogen to 37 &#176;C water bath was applied for the freeze-...

Watch this Scientific Journal Video about Development and Characterization of Decellularized Lung Extracellular Matrix Hydrogels at JoVE.com

The protocol elucidates two distinct decellularization methodologies applied to native bovine pulmonary tissues, providing a comprehensive account of their respective characterizations.

The use of extracellular matrix (ECM)-derived hydrogels in tissue engineering has become increasingly popular, as they can mimic cells&#39; natural ...