Name: culture and imaging of ex vivo organotypic pseudomyxoma peritonei tumor slices from resected human tumor specimens
Uploaded: 2022-12-09T13:55:00
Description: Watch this Scientific Journal Video about Culture and Imaging of Ex Vivo Organotypic Pseudomyxoma Peritonei Tumor Slices from Resected Human Tumor Specimens at JoVE.com

The authors would like to thank Kersi Pestonjamasp from the Moores Cancer Center imaging core facility for help with the microscopes UCSD Specialized Cancer Support Center P30 grant 2P30CA023100. This work was additionally supported by a JoVE publication grant (JRW), as well as generous gifts from the estate of Elisabeth and Ad Creemers, the Euske Family Foundation, the Gastrointestinal Cancer Research Fund, and the Peritoneal Metastasis Research Fund (AML).

The deidentification and acquisition of all tissues were performed under an IRB-approved protocol at the University of California, San Diego.
1. Preparation of human PMP tissues for tissue processing and culture
<ol>
	<li>Transport of tumor tissues and microdissection
	<ol>
		<li>Prepare the transport and culture media: complete 10% (v/v) Dulbecco&#39;s Modified Eagle Media (DMEM), 10% FBS, 2 mM L-Glutamine, 1% Penicillin/Streptomycin (Pen Strep).</li>
		<li>Upon tissue arri...

<ol>
	<li>Bevan, K. E., Mohamed, F., Moran, B. J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Pseudomyxoma+peritonei.">Pseudomyxoma peritonei.</a> World Journal of Gastrointestinal Oncology. 2 (1), 44-50 (2010).</li><li>Votanopoulos, K. I., 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=Appendiceal+cancer+patient-specific+tumor+organoid+model+for+predicting+chemotherapy+efficacy+prior+to+initiation+of+treatment:+A+feasibility+study.">Appendiceal cancer patient-specific tumor organoid model for predicting chemotherapy efficacy prior to initiation of treatment: A feasibility study.</a> Annals of Surgical Oncology. 26 (1), 139-147 (2019).</li><li>Holliday, D. L., 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=The+practicalities+of+using+tissue+slices+as+preclinical+organotypic+breast+cancer+models.">The practicalities of using tissue slices as preclinical organotypic breast cancer models.</a> Journal of Clinical Pathology. 66 (3), 253-255 (2013).</li><li>Koerfer, J., 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=Organotypic+slice+cultures+of+human+gastric+and+esophagogastric+junction+cancer.">Organotypic slice cultures of human gastric and esophagogastric junction cancer.</a> Cancer Medicine. 5 (7), 1444-1453 (2016).</li><li>Misra, S., 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=Ex+vivo+organotypic+culture+system+of+precision-cut+slices+of+human+pancreatic+ductal+adenocarcinoma.">Ex vivo organotypic culture system of precision-cut slices of human pancreatic ductal adenocarcinoma.</a> Scientific Reports. 9 (1), 2133 (2019).</li><li>Ohnishi, T., Matsumura, H., Izumoto, S., Hiraga, S., Hayakawa, T. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=A+novel+model+of+glioma+cell+invasion+using+organotypic+brain+slice+culture.">A novel model of glioma cell invasion using organotypic brain slice culture.</a> Cancer Research. 58 (14), 2935-2940 (1998).</li><li>Seidman, J. D., Elsayed, A. M., Sobin, L. H., Tavassoli, F. A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Association+of+mucinous+tumors+of+the+ovary+and+appendix.+A+clinicopathologic+study+of+25+cases.">Association of mucinous tumors of the ovary and appendix. A clinicopathologic study of 25 cases.</a> The Amerian Journal of Surgical Pathology. 17 (1), 22-34 (1993).</li><li>Mizuta, 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=Pseudomyxoma+peritonei+accompanied+by+intraductal+papillary+mucinous+neoplasm+of+the+pancreas.">Pseudomyxoma peritonei accompanied by intraductal papillary mucinous neoplasm of the pancreas.</a> Pancreatology. 5 (4-5), 470-474 (2005).</li><li>Gong, Y., Wang, X., Zhu, Z. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Pseudomyxoma+peritonei+originating+from+transverse+colon+mucinous+adenocarcinoma:+A+case+report+and+literature+review.">Pseudomyxoma peritonei originating from transverse colon mucinous adenocarcinoma: A case report and literature review.</a> Gastroenterology Research and Practice. 2020, 5826214 (2020).</li><li>Fleten, K. G., 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=Experimental+treatment+of+mucinous+peritoneal+metastases+using+patient-derived+xenograft+models.">Experimental treatment of mucinous peritoneal metastases using patient-derived xenograft models.</a> Translational Oncology. 13 (8), 100793 (2020).</li><li>Kuracha, M. R., Thomas, P., Loggie, B. W., Govindarajan, V. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Patient-derived+xenograft+mouse+models+of+pseudomyxoma+peritonei+recapitulate+the+human+inflammatory+tumor+microenvironment.">Patient-derived xenograft mouse models of pseudomyxoma peritonei recapitulate the human inflammatory tumor microenvironment.</a> Cancer Medicine. 5 (4), 711-719 (2016).</li><li>Jiang, 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=Long-lived+pancreatic+ductal+adenocarcinoma+slice+cultures+enable+precise+study+of+the+immune+microenvironment.">Long-lived pancreatic ductal adenocarcinoma slice cultures enable precise study of the immune microenvironment.</a> Oncoimmunology. 6 (7), 1333210 (2017).</li><li>Sundstrom, L., Morrison, B., Bradley, M., Pringle, A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Organotypic+cultures+as+tools+for+functional+screening+in+the+CNS.">Organotypic cultures as tools for functional screening in the CNS.</a> Drug Discovery Today. 10 (14), 993-1000 (2005).</li><li>Liu, L., Yu, L., Li, Z., Li, W., Huang, W. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Patient-derived+organoid+(PDO)+platforms+to+facilitate+clinical+decision+making.">Patient-derived organoid (PDO) platforms to facilitate clinical decision making.</a> Journal of Translational Medicine. 19 (1), 40 (2021).</li><li>Croft, C. L., Futch, H. S., Moore, B. D., Golde, T. E. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Organotypic+brain+slice+cultures+to+model+neurodegenerative+proteinopathies.">Organotypic brain slice cultures to model neurodegenerative proteinopathies.</a> Molecular Neurodegeneration. 14 (1), 45 (2019).</li><li>Carr, N. J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=New+insights+in+the+pathology+of+peritoneal+surface+malignancy.">New insights in the pathology of peritoneal surface malignancy.</a> Journal of Gastrointestinal Oncology. 12, 216-229 (2021).</li><li>Votanopoulos, K. I., 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=Outcomes+of+repeat+cytoreductive+surgery+with+hyperthermic+intraperitoneal+chemotherapy+for+the+treatment+of+peritoneal+surface+malignancy.">Outcomes of repeat cytoreductive surgery with hyperthermic intraperitoneal chemotherapy for the treatment of peritoneal surface malignancy.</a> Journal of the American College of Surgeons. 215 (3), 412-417 (2012).</li><li>Weitz, J., 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=An+ex-vivo+organotypic+culture+platform+for+functional+interrogation+of+human+appendiceal+cancer+reveals+a+prominent+and+heterogenous+immunological+landscape.">An ex-vivo organotypic culture platform for functional interrogation of human appendiceal cancer reveals a prominent and heterogenous immunological landscape.</a> Clinical Cancer Research. 28 (21), 4793-4806 (2022).</li><li>Pitoulis, F. G., Watson, S. A., Perbellini, F., Terracciano, C. M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Myocardial+slices+come+to+age:+an+intermediate+complexity+in+vitro+cardiac+model+for+translational+research.">Myocardial slices come to age: an intermediate complexity in vitro cardiac model for translational research.</a> Cardiovascular Research. 116 (7), 1275-1287 (2020).</li><li>Habeler, W., Peschanski, M., Monville, C. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Organotypic+heart+slices+for+cell+transplantation+and+physiological+studies.">Organotypic heart slices for cell transplantation and physiological studies.</a> Organogenesis. 5 (2), 62-66 (2009).</li></ol>

This manuscript describes a technique that can be used to culture, interrogate, and analyze human pseudomyxoma peritonei (PMP) tumor specimens. We have utilized numerous downstream functional assays to interrogate the tumor immune microenvironment and a platform for bench-to-bedside testing.
While the method is highly efficient in our hands, it will require some practice to cut tumor specimens using a vibratome. Namely, we encountered problems that were due to highly mucinous samples, as well ...

Pseudomyxoma peritonei (PMP) is rare syndrome with an incidence rate of 1 per million people per year1. Most PMP cases are caused by metastases from appendiceal neoplasms. Given that mice do not have a human-like appendix, modeling this type of cancer remains extremely challenging. While the primary disease is often curable by surgical resection, treatment options for metastatic disease are limited. Therefore, the rationale for developing this novel organotypic slice model is to study the pathobiology of PMP. To date, there are no appendiceal organoid models that can be perpetually cultured; however, a recent model was shown to be useful for the pharmacological testing of therapeutic agents and immunotherapy2. As such, we have adapted an organotypic slice culture system, which has been used in other types of human cancers, such as brain, breast, pancreas, lung, ovarian, and others3,4,5,6.
In addition to appendiceal neoplasms, PMP occasionally results from other tumor types, including ovarian cancers7, and in rare circumstances, intraductal papillary mucinous neoplasms8 and colon cancer9. Additionally, these tumors tend to grow slowly, with poor engraft rates in patient-derived xenograft (PDX) models10,11. Given these challenges, there is an unmet need to develop models to study this disease to begin to understand the pathobiology of PMP, and how these cancer cells: are recruited to the peritoneal surfaces, proliferate, and escape immune surveillance.
While cut from the systemic vascular circulation, tumor slices do contain cellular and acellular components, including the extracellular matrix, stromal cells, immune cells, cancer cells, endothelial cells, and nerves. This semi-intact microenvironment allows for the functional investigation of these cell types, which is uniquely advantageous compared to 3D organoid cultures, which consist only of cancer cells12. While organotypic slice cultures are advantageous in some respects, they are also inherently a low-throughput-based approach, compared to 3D organoids, which can be expanded, and are suitable for multiplexed investigational therapeutic drug screening13,14,15. In the case of PMP, there have been no reports documenting reliable establishment and perpetual passaging of PMP-derived organoids16. This is likely due to the slow growing nature of PMP-derived tumor cells, as well as the low number of malignant epithelial cells found within these mucinous tumors. Given the need to develop models to study PMP, organotypic slices are uniquely suited to study this disease. We present a protocol for preparing, imaging, and analyzing PMP from human specimens.

<table border="1" fo:keep-together.within-page="1" fo:keep-with-next.within-page="always">
	<tbody>
		<tr>
			<td>1 M CaCl2 solution</td>
			<td>Sigma</td>
			<td>21115</td>
			<td></td>
		</tr>
		<tr>
			<td>1 M HEPES solution</td>
			<td>Sigma</td>
			<td>H0887</td>
			<td></td>
		</tr>
		<tr>
			<td>1 M MgCl2 solution&#160;</td>
			<td>Sigma</td>
			<td>M1028</td>
			<td></td>
		</tr>
		<tr>
			<td>100 micron filter</td>
			<td>ThermoFisher</td>
			<td>22-363-549</td>
			<td></td>
		</tr>
		<tr>
			<td>22 x 40 glass coverslips</td>
			<td>Daiggerbrand</td>
			<td>G15972H</td>
			<td></td>
		</tr>
		<tr>
			<td>3 M KCl solution</td>
			<td>Sigma</td>
			<td>60135</td>
			<td></td>
		</tr>
		<tr>
			<td>5 M NaCl solution</td>
			<td>Sigma</td>
			<td>S5150</td>
			<td></td>
		</tr>
		<tr>
			<td>ATP&#947;S&#160;</td>
			<td>Tocris&#160;</td>
			<td>4080</td>
			<td></td>
		</tr>
		<tr>
			<td>Bovine Serum Albumin</td>
			<td>Sigma</td>
			<td>A2153</td>
			<td></td>
		</tr>
		<tr>
			<td>Calcein-AM&#160;</td>
			<td>Invitrogen</td>
			<td>L3224</td>
			<td></td>
		</tr>
		<tr>
			<td>CD11b&#160;</td>
			<td>Biolegend</td>
			<td>101228</td>
			<td></td>
		</tr>
		<tr>
			<td>CD206&#160;</td>
			<td>Biolegend</td>
			<td>321140</td>
			<td></td>
		</tr>
		<tr>
			<td>CD3</td>
			<td>Biolegend</td>
			<td>555333</td>
			<td></td>
		</tr>
		<tr>
			<td>CD4&#160;</td>
			<td>Biolegend</td>
			<td>357410</td>
			<td></td>
		</tr>
		<tr>
			<td>CD45&#160;</td>
			<td>Biolegend</td>
			<td>304006</td>
			<td></td>
		</tr>
		<tr>
			<td>CD8&#160;</td>
			<td>Biolegend</td>
			<td>344721</td>
			<td></td>
		</tr>
		<tr>
			<td>CellTiter-Glo&#160;</td>
			<td>Promega</td>
			<td>G9681</td>
			<td></td>
		</tr>
		<tr>
			<td>DMEM&#160;</td>
			<td>Thermo Fisher</td>
			<td>11965084</td>
			<td></td>
		</tr>
		<tr>
			<td>DPBS&#160;</td>
			<td>Sigma Aldrich</td>
			<td>D8537</td>
			<td></td>
		</tr>
		<tr>
			<td>FBS, heat inactivated</td>
			<td>ThermoFisher</td>
			<td>16140071</td>
			<td></td>
		</tr>
		<tr>
			<td>Fc-block&#160;</td>
			<td>BD Biosciences</td>
			<td>564220</td>
			<td></td>
		</tr>
		<tr>
			<td>Fluo-4</td>
			<td>Thermo Fisher</td>
			<td>F14201</td>
			<td></td>
		</tr>
		<tr>
			<td>Gentle Collagenase/Hyaluronidase&#160;</td>
			<td>Stem Cell</td>
			<td>7912</td>
			<td></td>
		</tr>
		<tr>
			<td>Imaging Chamber</td>
			<td>Warner Instruments</td>
			<td>RC-26</td>
			<td></td>
		</tr>
		<tr>
			<td>Imaging Chamber Platform</td>
			<td>Warner Instruments</td>
			<td>PH-1</td>
			<td></td>
		</tr>
		<tr>
			<td>LD-Blue&#160;</td>
			<td>Biolegend</td>
			<td>L23105</td>
			<td></td>
		</tr>
		<tr>
			<td>L-Glutamine 200 mM</td>
			<td>ThermoFisher</td>
			<td>25030081</td>
			<td></td>
		</tr>
		<tr>
			<td>LIVE/DEAD imaging dyes</td>
			<td>Thermofisher</td>
			<td>R37601</td>
			<td></td>
		</tr>
		<tr>
			<td>Nikon Ti microscope&#160;</td>
			<td>Nikon</td>
			<td></td>
			<td>Includes: A1R hybrid confocal scanner including a high-resolution (4096x4096) scanner, LU4 four-laser AOTF unit with 405, 488, 561, and 647 lasers, Plan Apo 10 (NA 0.8), 20X (NA 0.9) dry objectives.&#160;</td>
		</tr>
		<tr>
			<td>Peristaltic pump&#160;</td>
			<td>Isamtec</td>
			<td>ISM832C</td>
			<td></td>
		</tr>
		<tr>
			<td>Propidium Iodide</td>
			<td>Invitrogen</td>
			<td>L3224</td>
			<td></td>
		</tr>
		<tr>
			<td>Vacuum silicone grease</td>
			<td>Sigma</td>
			<td>Z273554-1EA</td>
			<td></td>
		</tr>
	</tbody>
</table>

culture and imaging of ex vivo organotypic pseudomyxoma peritonei tumor slices from resected human tumor specimens

Pseudomyxoma peritonei (PMP) is a rare condition that results from the dissemination of a mucinous primary tumor and the resultant accumulation of mucin-secreting tumor cells in the peritoneal cavity. PMP can arise from various types of cancers, including appendiceal, ovarian, and colorectal, though appendiceal neoplasms are by far the most common etiology. PMP is challenging to study due to its (1) rarity, (2) limited murine models, and (3) mucinous, acellular histology. The method presented here allows real-time visualization and interrogation of these tumor types using patient-derived ex vivo organotypic slices in a preparation where the tumor microenvironment (TME) remains intact. In this protocol, we first describe the preparation of tumor slices using a vibratome and subsequent long-term culture. Second, we describe confocal imaging of tumor slices and how to monitor functional readouts of viability, calcium imaging, and local proliferation. In short, slices are loaded with imaging dyes and are placed in an imaging chamber that can be mounted onto a confocal microscope. Time-lapse videos and confocal images are used to assess the initial viability and cellular functionality. This procedure also explores translational cellular movement, and paracrine signaling interactions in the TME. Lastly, we describe a dissociation protocol for tumor slices to be used for flow cytometry analysis. Quantitative flow cytometry analysis can be used for bench-to-bedside therapeutic testing to determine changes occurring within the immune landscape and epithelial cell content.

In short, human tumor specimens from PMP are obtained under an IRB-approved protocol. The tissue is prepared, micro-dissected, and solidified in an agarose mold to be cut using a vibratome (Figure 1A; Video 1). Once cut, tissue slices are placed and cultured on permeable insert membranes (Figure 1B), which can be utilized for imaging assays in situ, as well as for cellular and functional interrogation using flow cytometry, confocal imag...

Watch this Scientific Journal Video about Culture and Imaging of Ex Vivo Organotypic Pseudomyxoma Peritonei Tumor Slices from Resected Human Tumor Specimens at JoVE.com

Culture and Imaging of Ex Vivo Organotypic Pseudomyxoma Peritonei Tumor Slices from Resected Human Tumor Specimens

We describe a protocol for the production, culture, and visualization of human cancers, which have metastasized to the peritoneal surfaces. Resected tumor specimens are cut using a vibratome and cultured on permeable inserts for increased oxygenation and viability, followed by imaging and downstream analyses using confocal microscopy and flow cytometry.

Culture and Imaging of Ex Vivo Organotypic Pseudomyxoma Peritonei Tumor Slices from Resected Human Tumor Specimens

Pseudomyxoma peritonei (PMP) is a rare condition that results from the dissemination of a mucinous primary tumor and the resultant accumulation of ...

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