Smooth muscle cell injury after cryopreservation of human thoracic aortas
The cryopreservation protocol we use for arterial reconstructive surgery has been studied to evaluate smooth muscle cell (SMC) structural integrity and viability before implantation. Samples of human thoracic aortas (HTA) were harvested from five multi-organ donors. Sampling included unfrozen and cr...
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| Veröffentlicht in: | Cryobiology 2006-04, Vol.52 (2), p.309-316 |
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| creator | Pasquinelli, G. Foroni, L. Buzzi, M. Tazzari, P.L. Vaselli, C. Mirelli, M. Gargiulo, M. Conte, R. Stella, A. |
| description | The cryopreservation protocol we use for arterial reconstructive surgery has been studied to evaluate smooth muscle cell (SMC) structural integrity and viability before implantation. Samples of human thoracic aortas (HTA) were harvested from five multi-organ donors. Sampling included unfrozen and cryopreserved specimens. Cryopreservation was performed using RPMI with human albumin and 10% Me
2SO in a controlled-rate freezing apparatus. Thawing was accomplished by submerging bags in a water bath (39
°C) followed by washings in cooled saline. In situ cell preservation as investigated by light and transmission electron microscopy showed that SMCs from cryopreserved HTA had nuclear and cytoplasmic changes. A TUNEL assay, performed to detect DNA fragmentation in situ, showed increased SMC nuclear positivity in cryopreserved HTA when compared to unfrozen samples. 7-AAD flow cytometry assay of cells derived from cryopreserved HTA showed that an average of 49
±
16% cells were unlabeled after cryopreservation. Organ cultures aimed to study cell ability to recover cryopreservation damage showed a decreasing number of SMCs from day 4 to day 15 in cryopreserved HTA. In conclusion, the cryopreservation protocol applied in this study induces irreversible damage of a significant fraction of arterial SMCs. |
| doi_str_mv | 10.1016/j.cryobiol.2005.12.004 |
| format | Article |
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2SO in a controlled-rate freezing apparatus. Thawing was accomplished by submerging bags in a water bath (39
°C) followed by washings in cooled saline. In situ cell preservation as investigated by light and transmission electron microscopy showed that SMCs from cryopreserved HTA had nuclear and cytoplasmic changes. A TUNEL assay, performed to detect DNA fragmentation in situ, showed increased SMC nuclear positivity in cryopreserved HTA when compared to unfrozen samples. 7-AAD flow cytometry assay of cells derived from cryopreserved HTA showed that an average of 49
±
16% cells were unlabeled after cryopreservation. Organ cultures aimed to study cell ability to recover cryopreservation damage showed a decreasing number of SMCs from day 4 to day 15 in cryopreserved HTA. In conclusion, the cryopreservation protocol applied in this study induces irreversible damage of a significant fraction of arterial SMCs.</description><identifier>ISSN: 0011-2240</identifier><identifier>EISSN: 1090-2392</identifier><identifier>DOI: 10.1016/j.cryobiol.2005.12.004</identifier><identifier>PMID: 16458877</identifier><language>eng</language><publisher>Netherlands: Elsevier Inc</publisher><subject>Aorta, Thoracic - injuries ; Arterial banking ; Cryopreservation ; Cryoprotective Agents - chemistry ; Flow cytometry ; Human aortic homografts ; Humans ; Immunohistochemistry ; In Situ Nick-End Labeling ; Microscopy, Electron, Transmission ; Myocytes, Smooth Muscle - pathology ; Organ culture ; Organ Culture Techniques ; Organ Preservation ; Smooth muscle cell injury ; Tissue Survival ; Transmission electron microscopy ; TUNEL</subject><ispartof>Cryobiology, 2006-04, Vol.52 (2), p.309-316</ispartof><rights>2005 Elsevier Inc.</rights><lds50>peer_reviewed</lds50><woscitedreferencessubscribed>false</woscitedreferencessubscribed><citedby>FETCH-LOGICAL-c397t-1764b02d79953b739f045a2b13e1973fb0b4cf3c443b1e060a114d26175c5d9b3</citedby><cites>FETCH-LOGICAL-c397t-1764b02d79953b739f045a2b13e1973fb0b4cf3c443b1e060a114d26175c5d9b3</cites></display><links><openurl>$$Topenurl_article</openurl><openurlfulltext>$$Topenurlfull_article</openurlfulltext><thumbnail>$$Tsyndetics_thumb_exl</thumbnail><linktohtml>$$Uhttps://www.sciencedirect.com/science/article/pii/S0011224005001963$$EHTML$$P50$$Gelsevier$$H</linktohtml><link.rule.ids>314,776,780,3537,27901,27902,65306</link.rule.ids><backlink>$$Uhttps://www.ncbi.nlm.nih.gov/pubmed/16458877$$D View this record in MEDLINE/PubMed$$Hfree_for_read</backlink></links><search><creatorcontrib>Pasquinelli, G.</creatorcontrib><creatorcontrib>Foroni, L.</creatorcontrib><creatorcontrib>Buzzi, M.</creatorcontrib><creatorcontrib>Tazzari, P.L.</creatorcontrib><creatorcontrib>Vaselli, C.</creatorcontrib><creatorcontrib>Mirelli, M.</creatorcontrib><creatorcontrib>Gargiulo, M.</creatorcontrib><creatorcontrib>Conte, R.</creatorcontrib><creatorcontrib>Stella, A.</creatorcontrib><title>Smooth muscle cell injury after cryopreservation of human thoracic aortas</title><title>Cryobiology</title><addtitle>Cryobiology</addtitle><description>The cryopreservation protocol we use for arterial reconstructive surgery has been studied to evaluate smooth muscle cell (SMC) structural integrity and viability before implantation. Samples of human thoracic aortas (HTA) were harvested from five multi-organ donors. Sampling included unfrozen and cryopreserved specimens. Cryopreservation was performed using RPMI with human albumin and 10% Me
2SO in a controlled-rate freezing apparatus. Thawing was accomplished by submerging bags in a water bath (39
°C) followed by washings in cooled saline. In situ cell preservation as investigated by light and transmission electron microscopy showed that SMCs from cryopreserved HTA had nuclear and cytoplasmic changes. A TUNEL assay, performed to detect DNA fragmentation in situ, showed increased SMC nuclear positivity in cryopreserved HTA when compared to unfrozen samples. 7-AAD flow cytometry assay of cells derived from cryopreserved HTA showed that an average of 49
±
16% cells were unlabeled after cryopreservation. Organ cultures aimed to study cell ability to recover cryopreservation damage showed a decreasing number of SMCs from day 4 to day 15 in cryopreserved HTA. In conclusion, the cryopreservation protocol applied in this study induces irreversible damage of a significant fraction of arterial SMCs.</description><subject>Aorta, Thoracic - injuries</subject><subject>Arterial banking</subject><subject>Cryopreservation</subject><subject>Cryoprotective Agents - chemistry</subject><subject>Flow cytometry</subject><subject>Human aortic homografts</subject><subject>Humans</subject><subject>Immunohistochemistry</subject><subject>In Situ Nick-End Labeling</subject><subject>Microscopy, Electron, Transmission</subject><subject>Myocytes, Smooth Muscle - pathology</subject><subject>Organ culture</subject><subject>Organ Culture Techniques</subject><subject>Organ Preservation</subject><subject>Smooth muscle cell injury</subject><subject>Tissue Survival</subject><subject>Transmission electron microscopy</subject><subject>TUNEL</subject><issn>0011-2240</issn><issn>1090-2392</issn><fulltext>true</fulltext><rsrctype>article</rsrctype><creationdate>2006</creationdate><recordtype>article</recordtype><sourceid>EIF</sourceid><recordid>eNqFkE1v1DAURa0KRIfCX6i8Ypfwnu3Ykx2ooh9SJRbA2rKdF41HSTy1k0rz78loBrHs6m3OvffpMHaLUCOg_rqvQz4mH9NQC4CmRlEDqCu2QWihErIV79gGALESQsE1-1jKHgC0keoDu0atmu3WmA17-jWmNO_4uJQwEA80DDxO-yUfuetnyvw0c8hUKL-6OaaJp57vltFNfN6l7EIM3KU8u_KJve_dUOjz5d6wP_c_ft89Vs8_H57uvj9XQbZmrtBo5UF0pm0b6Y1se1CNEx4lYWtk78Gr0MuglPRIoMEhqk5oNE1outbLG_bl3HvI6WWhMtsxltPfbqK0FKuNkcZo-SaIBqHZol5BfQZDTqVk6u0hx9Hlo0WwJ9t2b__ZtifbFoVdba_B28vC4kfq_scuelfg2xmgVchrpGxLiDQF6mKmMNsuxbc2_gK6s5QK</recordid><startdate>20060401</startdate><enddate>20060401</enddate><creator>Pasquinelli, G.</creator><creator>Foroni, L.</creator><creator>Buzzi, M.</creator><creator>Tazzari, P.L.</creator><creator>Vaselli, C.</creator><creator>Mirelli, M.</creator><creator>Gargiulo, M.</creator><creator>Conte, R.</creator><creator>Stella, A.</creator><general>Elsevier Inc</general><scope>CGR</scope><scope>CUY</scope><scope>CVF</scope><scope>ECM</scope><scope>EIF</scope><scope>NPM</scope><scope>AAYXX</scope><scope>CITATION</scope><scope>7QO</scope><scope>8FD</scope><scope>FR3</scope><scope>P64</scope><scope>7X8</scope></search><sort><creationdate>20060401</creationdate><title>Smooth muscle cell injury after cryopreservation of human thoracic aortas</title><author>Pasquinelli, G. ; Foroni, L. ; Buzzi, M. ; Tazzari, P.L. ; Vaselli, C. ; Mirelli, M. ; Gargiulo, M. ; Conte, R. ; Stella, A.</author></sort><facets><frbrtype>5</frbrtype><frbrgroupid>cdi_FETCH-LOGICAL-c397t-1764b02d79953b739f045a2b13e1973fb0b4cf3c443b1e060a114d26175c5d9b3</frbrgroupid><rsrctype>articles</rsrctype><prefilter>articles</prefilter><language>eng</language><creationdate>2006</creationdate><topic>Aorta, Thoracic - injuries</topic><topic>Arterial banking</topic><topic>Cryopreservation</topic><topic>Cryoprotective Agents - chemistry</topic><topic>Flow cytometry</topic><topic>Human aortic homografts</topic><topic>Humans</topic><topic>Immunohistochemistry</topic><topic>In Situ Nick-End Labeling</topic><topic>Microscopy, Electron, Transmission</topic><topic>Myocytes, Smooth Muscle - pathology</topic><topic>Organ culture</topic><topic>Organ Culture Techniques</topic><topic>Organ Preservation</topic><topic>Smooth muscle cell injury</topic><topic>Tissue Survival</topic><topic>Transmission electron microscopy</topic><topic>TUNEL</topic><toplevel>peer_reviewed</toplevel><toplevel>online_resources</toplevel><creatorcontrib>Pasquinelli, G.</creatorcontrib><creatorcontrib>Foroni, L.</creatorcontrib><creatorcontrib>Buzzi, M.</creatorcontrib><creatorcontrib>Tazzari, P.L.</creatorcontrib><creatorcontrib>Vaselli, C.</creatorcontrib><creatorcontrib>Mirelli, M.</creatorcontrib><creatorcontrib>Gargiulo, M.</creatorcontrib><creatorcontrib>Conte, R.</creatorcontrib><creatorcontrib>Stella, A.</creatorcontrib><collection>Medline</collection><collection>MEDLINE</collection><collection>MEDLINE (Ovid)</collection><collection>MEDLINE</collection><collection>MEDLINE</collection><collection>PubMed</collection><collection>CrossRef</collection><collection>Biotechnology Research Abstracts</collection><collection>Technology Research Database</collection><collection>Engineering Research Database</collection><collection>Biotechnology and BioEngineering Abstracts</collection><collection>MEDLINE - Academic</collection><jtitle>Cryobiology</jtitle></facets><delivery><delcategory>Remote Search Resource</delcategory><fulltext>fulltext</fulltext></delivery><addata><au>Pasquinelli, G.</au><au>Foroni, L.</au><au>Buzzi, M.</au><au>Tazzari, P.L.</au><au>Vaselli, C.</au><au>Mirelli, M.</au><au>Gargiulo, M.</au><au>Conte, R.</au><au>Stella, A.</au><format>journal</format><genre>article</genre><ristype>JOUR</ristype><atitle>Smooth muscle cell injury after cryopreservation of human thoracic aortas</atitle><jtitle>Cryobiology</jtitle><addtitle>Cryobiology</addtitle><date>2006-04-01</date><risdate>2006</risdate><volume>52</volume><issue>2</issue><spage>309</spage><epage>316</epage><pages>309-316</pages><issn>0011-2240</issn><eissn>1090-2392</eissn><abstract>The cryopreservation protocol we use for arterial reconstructive surgery has been studied to evaluate smooth muscle cell (SMC) structural integrity and viability before implantation. Samples of human thoracic aortas (HTA) were harvested from five multi-organ donors. Sampling included unfrozen and cryopreserved specimens. Cryopreservation was performed using RPMI with human albumin and 10% Me
2SO in a controlled-rate freezing apparatus. Thawing was accomplished by submerging bags in a water bath (39
°C) followed by washings in cooled saline. In situ cell preservation as investigated by light and transmission electron microscopy showed that SMCs from cryopreserved HTA had nuclear and cytoplasmic changes. A TUNEL assay, performed to detect DNA fragmentation in situ, showed increased SMC nuclear positivity in cryopreserved HTA when compared to unfrozen samples. 7-AAD flow cytometry assay of cells derived from cryopreserved HTA showed that an average of 49
±
16% cells were unlabeled after cryopreservation. Organ cultures aimed to study cell ability to recover cryopreservation damage showed a decreasing number of SMCs from day 4 to day 15 in cryopreserved HTA. In conclusion, the cryopreservation protocol applied in this study induces irreversible damage of a significant fraction of arterial SMCs.</abstract><cop>Netherlands</cop><pub>Elsevier Inc</pub><pmid>16458877</pmid><doi>10.1016/j.cryobiol.2005.12.004</doi><tpages>8</tpages></addata></record> |
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| subjects | Aorta, Thoracic - injuries Arterial banking Cryopreservation Cryoprotective Agents - chemistry Flow cytometry Human aortic homografts Humans Immunohistochemistry In Situ Nick-End Labeling Microscopy, Electron, Transmission Myocytes, Smooth Muscle - pathology Organ culture Organ Culture Techniques Organ Preservation Smooth muscle cell injury Tissue Survival Transmission electron microscopy TUNEL |
| title | Smooth muscle cell injury after cryopreservation of human thoracic aortas |
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