- Email: [email protected]
The grand challenges of organ banking: How the world is beginning to align to meet them
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Abstracts / Cryobiology 71 (2015) 537e573
S37. THE GRAND CHALLENGES OF ORGAN BANKING: HOW THE WORLD IS BEGINNING TO ALIGN TO MEET THEM
S39. APPLICATION OF COMPUTED TOMOGRAPHY TO OVARIAN TISSUE CRYOPRESERVATION BY SLOW FREEZING
Sebastian Giwa 1, 2, Jedd Lewis 1, 3, Alessandro Tocchio 1, 3. 1 Organ Preservation Alliance, Moffett Field, California, USA; 2 Sylvatica Biotech, Inc., New York, New York, USA; 3 Stanford School of Medicine, Stanford University, Stanford, California, USA
Ariadna Corral, Marcin Balcerzyk, Angel Parrado, Christiani n Amorim, Marie-Madeleine Dolmans, Julio de la Fuente, Ramo Risco. University of Sevilla, Sevilla, Spain E-mail address: [email protected]
E-mail address: [email protected]
The Organ Preservation Alliance (a non-profit organization incubated at SU Labs, at NASA Research Park in Silicon Valley) is working to catalyze breakthroughs in complex tissue and organ banking to revolutionize transplant, trauma, and regenerative medicine. The organ shortage has grown steadily for decades in the U.S. and is even more severe in most other parts of the world. Poor donor utilization and high rates of organ discard remain major contributors to this shortage, fueled in many ways by the short preservation times of transplant organs and inability to adequately match donors and recipients. While advances in transplantation and bioengineering hold promise to reduce or eventually eliminate shortages, they have been met by slow progress in the preservation of large tissues. As regenerative medicine comes of age, tissue preservation has become a bottleneck. Cryopreservation holds the promise to meet this challenge and transform transplant medicine, resulting in fewer discards, better matching, fewer complications, and lower costs. It will allow for on-demand tissues for military and civilian trauma care as well as stockpiling for mass casualty events. It can enable new technologies and procedures, giving bioengineered tissues a shelf life, protecting fertility in cancer patients, facilitating new transplant immune tolerance protocols, and more. The Alliance recently, in partnership with the Society for Cryobiology, convened the first global Organ Banking Summit at Stanford University (with side-sessions at Lawrence Berkley National Laboratory and NASA Research Park), co-lead an NSF-funded Organ Banking & Bioengineering Roadmap Process, and convened a Washington D.C. Roadmap Workshop for federal agencies, leading scientists, foundations and industry - that was followed by a White House Roundtable Meeting. Funding: N/A Conflict of Interests: N/A S38. TROPHIC FACTORS OF FETAL ORIGIN IN PRESERVATION SOLUTION SUPPORTS ISOLATED LIVER DURING COLD STORAGE A. Yu Petrenko, D.V. Cherkashina, I.A. Sosimchik, O.A. Semenchenko, A. Yu Semenchenko. Institute for Problems of Cryobiology and Cryomedicine of the National Academy of Sciences of Ukraine, Kharkov, Ukraine E-mail address: [email protected]
Safe organ cold storage (CS) is an actual problem of cryobiology. It is promising to enrich the preservation solutions with a cocktail of growth factors and cytokines. Earlier we showed that animal pretreatment with trophic factors of fetal origin (TFFO) reduced liver damage after 24 hour CS and warm reperfusion (WR). The aim of this work is to study the effects of TFFO added to preservation solution on liver state after CS and WR. Rat fetal mesenchymal-mesodermal tissues (15-16 ED) were used as a source of TFFO. Rat livers were stored for 24 hours at 4 С without or with TFFO (20 mg of protein/ml), then reperfused during 60 min at 37 С. Liver CS and following WR led to uncoupling of oxidative phosphorylation, prooxidant/antioxidant disbalance, decrease of ATP level and bile production. Presence of TFFO prevented impairment of mitochondria function, promoting recovery of ATP content. ROS production was decreased after CS, TBARS level e after WR, LPO intensity was lower than in intact livers. Partial recovery of antioxidant enzyme activities was observed. TFFO presence maintained bile production rate and liver morphological structure similar to intact organs. Pronounced protective effect of TFFO on livers indicates the prospects of their application as compounds of medium for cold preservation. Funding: N/A Conflict of Interests: N/A
Ovarian tissue cryopreservation for women treated for cancer is in most cases the only way to preserve their fertility. The survival rate of the follicle population is still around a few percent in successful cases, mostly cryopreserved by slow freezing. Computed tomography (CT) has been applied to monitor cryoprotectant (CPA) concentration and ice formation inside ovarian tissue samples. The sulfur atom of the dimethyl sulfoxide (Me2SO) makes the X-ray attenuation proportional to concentration in this particular CPA. Human and animal ovarian tissue samples of an average size of 5x5x1 mm3 have been cryopreserved with slow freezing conventional protocols1 and analyzed with the CT at temperatures below -140 C, and at room temperature after rewarming and washing. Results show how tissue samples concentrated up to 80% v/v Me2SO, but in a non-homogeneous way. Moreover ice formation was observed in some parts of the tissues. CT technology is a very useful tool that can be used to assess the final CPA concentration and the ice formation in a cryopreserved sample, allowing tuning of the necessary parameters to achieve an ovarian tissue cryopreservation with a higher rate of survival. 1Dolmans MM et al, J Assist Reprod Genet 30 (2013), 305. Funding: N/A Conflict of Interests: N/A S40. CRYOPRESERVATION OF VASCULAR GRAFTS FOR CLINICAL USE e RETROSPECTIVE ANALYSIS OF PRE-FREEZING FACTORS WITH POTENTIAL IMPACT ON THE QUALITY AND SAFETY OF VASCULAR TRANSPLANTATIONS a dler 4, R. ri cek 2, L. Janousek 3, L. Dvora cek 4, P. St P. Me cka 1, M. Spa 5 6 1 1 rba 1, B. Vlachovský , I. Gunka , B. Honegrova , D. Brandejs , L. Ste e pa nov til 8, R. Staffa 5, P. Ne mec 9, J. a 7, P. Navra Voxov a 7, V. St lov e, Czech Lindner 2. 1 Tissue Bank, University Hospital Hradec Kra Prepublic; 2 II.nd. Dept. of Surgery e Cardiovascular Surgery of the 1st Faculty of Medicine Charles University in Prague and General University Hospital, Prague, Czech Republic; 3 Dept. of Transplantation Surgery, Inst. of Clinical and Experimental Medicine, Prague, Czech Republic; 4 Dept. of Vascular Surgery, Hospital Na Homolce, Prague, Czech Republic; 5 II.nd. Dept. of Surgery of the Medical Faculty, Masaryk University and St. Anne’s University Hospital, Brno, Czech Republic; 6 Dept. of Surgery of the Charles lov University in Prague, Medical Faculty Hradec Kra e and University lov Hospital Hradec Kra e, Czech Republic; 7 Dept. of Clinical Microbiology lov Charles University in Prague, Medical Faculty Hradec Kra e and lov University Hospital Hradec Kra e, Czech Republic; 8 Regional Transplantation Centre, Dept. of Urology, University Hospital Hradec lov Kra e, Czech Republic; 9 Centre of Cardiovascular and Transplantation Surgery, Brno, Czech Republic E-mail address: [email protected]
Aim: To analyse risk factors, such as: donor’s age (DA); time between tissue harvest and mixing with the cryoprotectant (TP); exposure to the cryoprotectant before freezing (CPAE); and the initial contamination rate (MCR). Methods: The data of 30 arterial (AG) and 30 venous grafts (VG) collected during the multiple organ harvests were analyzed. Only grafts meeting the criteria of release for clinical application including the proof of sterility at output control were included. The grafts were transported in the precooled organ preservation solution containing gentamycin. After input control and decontamination procedure each graft was put into a double plastic bag filled with 50ml of pre-cooled 10% hydroxyethlystarch solution and an equal volume of pre-cooled 20% (v/v) dimethylsulphoxide solution was added. Controlled freezing followed.
Abstracts / Cryobiology 71 (2015) 537e573
S37. THE GRAND CHALLENGES OF ORGAN BANKING: HOW THE WORLD IS BEGINNING TO ALIGN TO MEET THEM
S39. APPLICATION OF COMPUTED TOMOGRAPHY TO OVARIAN TISSUE CRYOPRESERVATION BY SLOW FREEZING
Sebastian Giwa 1, 2, Jedd Lewis 1, 3, Alessandro Tocchio 1, 3. 1 Organ Preservation Alliance, Moffett Field, California, USA; 2 Sylvatica Biotech, Inc., New York, New York, USA; 3 Stanford School of Medicine, Stanford University, Stanford, California, USA
Ariadna Corral, Marcin Balcerzyk, Angel Parrado, Christiani n Amorim, Marie-Madeleine Dolmans, Julio de la Fuente, Ramo Risco. University of Sevilla, Sevilla, Spain E-mail address: [email protected]
E-mail address: [email protected]
The Organ Preservation Alliance (a non-profit organization incubated at SU Labs, at NASA Research Park in Silicon Valley) is working to catalyze breakthroughs in complex tissue and organ banking to revolutionize transplant, trauma, and regenerative medicine. The organ shortage has grown steadily for decades in the U.S. and is even more severe in most other parts of the world. Poor donor utilization and high rates of organ discard remain major contributors to this shortage, fueled in many ways by the short preservation times of transplant organs and inability to adequately match donors and recipients. While advances in transplantation and bioengineering hold promise to reduce or eventually eliminate shortages, they have been met by slow progress in the preservation of large tissues. As regenerative medicine comes of age, tissue preservation has become a bottleneck. Cryopreservation holds the promise to meet this challenge and transform transplant medicine, resulting in fewer discards, better matching, fewer complications, and lower costs. It will allow for on-demand tissues for military and civilian trauma care as well as stockpiling for mass casualty events. It can enable new technologies and procedures, giving bioengineered tissues a shelf life, protecting fertility in cancer patients, facilitating new transplant immune tolerance protocols, and more. The Alliance recently, in partnership with the Society for Cryobiology, convened the first global Organ Banking Summit at Stanford University (with side-sessions at Lawrence Berkley National Laboratory and NASA Research Park), co-lead an NSF-funded Organ Banking & Bioengineering Roadmap Process, and convened a Washington D.C. Roadmap Workshop for federal agencies, leading scientists, foundations and industry - that was followed by a White House Roundtable Meeting. Funding: N/A Conflict of Interests: N/A S38. TROPHIC FACTORS OF FETAL ORIGIN IN PRESERVATION SOLUTION SUPPORTS ISOLATED LIVER DURING COLD STORAGE A. Yu Petrenko, D.V. Cherkashina, I.A. Sosimchik, O.A. Semenchenko, A. Yu Semenchenko. Institute for Problems of Cryobiology and Cryomedicine of the National Academy of Sciences of Ukraine, Kharkov, Ukraine E-mail address: [email protected]
Safe organ cold storage (CS) is an actual problem of cryobiology. It is promising to enrich the preservation solutions with a cocktail of growth factors and cytokines. Earlier we showed that animal pretreatment with trophic factors of fetal origin (TFFO) reduced liver damage after 24 hour CS and warm reperfusion (WR). The aim of this work is to study the effects of TFFO added to preservation solution on liver state after CS and WR. Rat fetal mesenchymal-mesodermal tissues (15-16 ED) were used as a source of TFFO. Rat livers were stored for 24 hours at 4 С without or with TFFO (20 mg of protein/ml), then reperfused during 60 min at 37 С. Liver CS and following WR led to uncoupling of oxidative phosphorylation, prooxidant/antioxidant disbalance, decrease of ATP level and bile production. Presence of TFFO prevented impairment of mitochondria function, promoting recovery of ATP content. ROS production was decreased after CS, TBARS level e after WR, LPO intensity was lower than in intact livers. Partial recovery of antioxidant enzyme activities was observed. TFFO presence maintained bile production rate and liver morphological structure similar to intact organs. Pronounced protective effect of TFFO on livers indicates the prospects of their application as compounds of medium for cold preservation. Funding: N/A Conflict of Interests: N/A
Ovarian tissue cryopreservation for women treated for cancer is in most cases the only way to preserve their fertility. The survival rate of the follicle population is still around a few percent in successful cases, mostly cryopreserved by slow freezing. Computed tomography (CT) has been applied to monitor cryoprotectant (CPA) concentration and ice formation inside ovarian tissue samples. The sulfur atom of the dimethyl sulfoxide (Me2SO) makes the X-ray attenuation proportional to concentration in this particular CPA. Human and animal ovarian tissue samples of an average size of 5x5x1 mm3 have been cryopreserved with slow freezing conventional protocols1 and analyzed with the CT at temperatures below -140 C, and at room temperature after rewarming and washing. Results show how tissue samples concentrated up to 80% v/v Me2SO, but in a non-homogeneous way. Moreover ice formation was observed in some parts of the tissues. CT technology is a very useful tool that can be used to assess the final CPA concentration and the ice formation in a cryopreserved sample, allowing tuning of the necessary parameters to achieve an ovarian tissue cryopreservation with a higher rate of survival. 1Dolmans MM et al, J Assist Reprod Genet 30 (2013), 305. Funding: N/A Conflict of Interests: N/A S40. CRYOPRESERVATION OF VASCULAR GRAFTS FOR CLINICAL USE e RETROSPECTIVE ANALYSIS OF PRE-FREEZING FACTORS WITH POTENTIAL IMPACT ON THE QUALITY AND SAFETY OF VASCULAR TRANSPLANTATIONS a dler 4, R. ri cek 2, L. Janousek 3, L. Dvora cek 4, P. St P. Me cka 1, M. Spa 5 6 1 1 rba 1, B. Vlachovský , I. Gunka , B. Honegrova , D. Brandejs , L. Ste e pa nov til 8, R. Staffa 5, P. Ne mec 9, J. a 7, P. Navra Voxov a 7, V. St lov e, Czech Lindner 2. 1 Tissue Bank, University Hospital Hradec Kra Prepublic; 2 II.nd. Dept. of Surgery e Cardiovascular Surgery of the 1st Faculty of Medicine Charles University in Prague and General University Hospital, Prague, Czech Republic; 3 Dept. of Transplantation Surgery, Inst. of Clinical and Experimental Medicine, Prague, Czech Republic; 4 Dept. of Vascular Surgery, Hospital Na Homolce, Prague, Czech Republic; 5 II.nd. Dept. of Surgery of the Medical Faculty, Masaryk University and St. Anne’s University Hospital, Brno, Czech Republic; 6 Dept. of Surgery of the Charles lov University in Prague, Medical Faculty Hradec Kra e and University lov Hospital Hradec Kra e, Czech Republic; 7 Dept. of Clinical Microbiology lov Charles University in Prague, Medical Faculty Hradec Kra e and lov University Hospital Hradec Kra e, Czech Republic; 8 Regional Transplantation Centre, Dept. of Urology, University Hospital Hradec lov Kra e, Czech Republic; 9 Centre of Cardiovascular and Transplantation Surgery, Brno, Czech Republic E-mail address: [email protected]
Aim: To analyse risk factors, such as: donor’s age (DA); time between tissue harvest and mixing with the cryoprotectant (TP); exposure to the cryoprotectant before freezing (CPAE); and the initial contamination rate (MCR). Methods: The data of 30 arterial (AG) and 30 venous grafts (VG) collected during the multiple organ harvests were analyzed. Only grafts meeting the criteria of release for clinical application including the proof of sterility at output control were included. The grafts were transported in the precooled organ preservation solution containing gentamycin. After input control and decontamination procedure each graft was put into a double plastic bag filled with 50ml of pre-cooled 10% hydroxyethlystarch solution and an equal volume of pre-cooled 20% (v/v) dimethylsulphoxide solution was added. Controlled freezing followed.