Hongyan Sun1,2, Ran Sun1, Miao Hao1, Yuqian Wang1, Xuewen Zhang3, Ya Liu2, Xianling Cong4. 1. Tissue Bank, China-Japan Union Hospital of Jilin University, Jilin, China. 2. Department of Nutrition and Food Hygiene, School of Public Health of Jilin University, Jilin, China. 3. Department of Surgery, China-Japan Union Hospital of Jilin University, Jilin, China. 4. Tissue Bank, China-Japan Union Hospital of Jilin University, Jilin, China. congxl888@hotmail.com.
Abstract
BACKGROUND: RNA degradation is a major problem in tissue banking, and the effects of the ex vivo ischemia time, storage time, and transport conditions on RNA integrity and gene expression have not been well understood. METHODS: A total of 100 fresh-frozen clear cell carcinoma tissues and matched normal tissues during five storage periods (≤6, 7-12, 13-18, 19-24, and 25-30 months) were chosen to detect RNA quality. At surgery, fresh kidney cancer tissues from five patients were cut into pieces and snap frozen. Additional fresh tissue pieces were (1) left at room temperature, (2) kept on ice, or (3) placed in normal saline before being snap frozen after 0.5, 1, 2, or 4 h. RNA integrity was determined by microchip electrophoresis, and gene expression was analyzed by real-time polymerase chain reaction. RESULTS: Altogether, 82 % of kidney cancer specimens banked using a standardized protocol yielded RNA with an RNA integrity number of ≥7 (7.77 ± 0.95, 7.87 ± 0.37, 7.15 ± 1.46, 8.10 ± 0.64, and 7.11 ± 1.08 during the five storage periods, respectively). RNA remained intact after 4 h on ice, whereas degradation was found in tissues left at room temperature or in saline. Expression of genes in certain functional pathways changed during storage under three conditions. CONCLUSIONS: More than 80 % of the banked kidney cancer biospecimens collected following a standardized protocol yielded high-quality RNA. Fresh human cancer tissue samples should be transported on ice before biobanking to avoid a major reduction in RNA quality. The presented data should be considered in attempts to further standardize tissue biospecimen collection and banking.
BACKGROUND: RNA degradation is a major problem in tissue banking, and the effects of the ex vivo ischemia time, storage time, and transport conditions on RNA integrity and gene expression have not been well understood. METHODS: A total of 100 fresh-frozen clear cell carcinoma tissues and matched normal tissues during five storage periods (≤6, 7-12, 13-18, 19-24, and 25-30 months) were chosen to detect RNA quality. At surgery, fresh kidney cancer tissues from five patients were cut into pieces and snap frozen. Additional fresh tissue pieces were (1) left at room temperature, (2) kept on ice, or (3) placed in normal saline before being snap frozen after 0.5, 1, 2, or 4 h. RNA integrity was determined by microchip electrophoresis, and gene expression was analyzed by real-time polymerase chain reaction. RESULTS: Altogether, 82 % of kidney cancer specimens banked using a standardized protocol yielded RNA with an RNA integrity number of ≥7 (7.77 ± 0.95, 7.87 ± 0.37, 7.15 ± 1.46, 8.10 ± 0.64, and 7.11 ± 1.08 during the five storage periods, respectively). RNA remained intact after 4 h on ice, whereas degradation was found in tissues left at room temperature or in saline. Expression of genes in certain functional pathways changed during storage under three conditions. CONCLUSIONS: More than 80 % of the banked kidney cancer biospecimens collected following a standardized protocol yielded high-quality RNA. Fresh humancancer tissue samples should be transported on ice before biobanking to avoid a major reduction in RNA quality. The presented data should be considered in attempts to further standardize tissue biospecimen collection and banking.
Authors: Douglas G Walker; Alexis M Whetzel; Geidy Serrano; Lucia I Sue; Lih-Fen Lue; Thomas G Beach Journal: Cell Tissue Bank Date: 2016-04-16 Impact factor: 1.522
Authors: Thai H Ho; Rafael Nunez Nateras; Huihuang Yan; Jin G Park; Sally Jensen; Chad Borges; Jeong Heon Lee; Mia D Champion; Raoul Tibes; Alan H Bryce; Estrella M Carballido; Mark A Todd; Richard W Joseph; William W Wong; Alexander S Parker; Melissa L Stanton; Erik P Castle Journal: PLoS One Date: 2015-07-16 Impact factor: 3.240
Authors: Andrea Cossarizza; Hyun-Dong Chang; Andreas Radbruch; Andreas Acs; Dieter Adam; Sabine Adam-Klages; William W Agace; Nima Aghaeepour; Mübeccel Akdis; Matthieu Allez; Larissa Nogueira Almeida; Giorgia Alvisi; Graham Anderson; Immanuel Andrä; Francesco Annunziato; Achille Anselmo; Petra Bacher; Cosima T Baldari; Sudipto Bari; Vincenzo Barnaba; Joana Barros-Martins; Luca Battistini; Wolfgang Bauer; Sabine Baumgart; Nicole Baumgarth; Dirk Baumjohann; Bianka Baying; Mary Bebawy; Burkhard Becher; Wolfgang Beisker; Vladimir Benes; Rudi Beyaert; Alfonso Blanco; Dominic A Boardman; Christian Bogdan; Jessica G Borger; Giovanna Borsellino; Philip E Boulais; Jolene A Bradford; Dirk Brenner; Ryan R Brinkman; Anna E S Brooks; Dirk H Busch; Martin Büscher; Timothy P Bushnell; Federica Calzetti; Garth Cameron; Ilenia Cammarata; Xuetao Cao; Susanna L Cardell; Stefano Casola; Marco A Cassatella; Andrea Cavani; Antonio Celada; Lucienne Chatenoud; Pratip K Chattopadhyay; Sue Chow; Eleni Christakou; Luka Čičin-Šain; Mario Clerici; Federico S Colombo; Laura Cook; Anne Cooke; Andrea M Cooper; Alexandra J Corbett; Antonio Cosma; Lorenzo Cosmi; Pierre G Coulie; Ana Cumano; Ljiljana Cvetkovic; Van Duc Dang; Chantip Dang-Heine; Martin S Davey; Derek Davies; Sara De Biasi; Genny Del Zotto; Gelo Victoriano Dela Cruz; Michael Delacher; Silvia Della Bella; Paolo Dellabona; Günnur Deniz; Mark Dessing; James P Di Santo; Andreas Diefenbach; Francesco Dieli; Andreas Dolf; Thomas Dörner; Regine J Dress; Diana Dudziak; Michael Dustin; Charles-Antoine Dutertre; Friederike Ebner; Sidonia B G Eckle; Matthias Edinger; Pascale Eede; Götz R A Ehrhardt; Marcus Eich; Pablo Engel; Britta Engelhardt; Anna Erdei; Charlotte Esser; Bart Everts; Maximilien Evrard; Christine S Falk; Todd A Fehniger; Mar Felipo-Benavent; Helen Ferry; Markus Feuerer; Andrew Filby; Kata Filkor; Simon Fillatreau; Marie Follo; Irmgard Förster; John Foster; Gemma A Foulds; Britta Frehse; Paul S Frenette; Stefan Frischbutter; Wolfgang Fritzsche; David W Galbraith; Anastasia Gangaev; Natalio Garbi; Brice Gaudilliere; Ricardo T Gazzinelli; Jens Geginat; Wilhelm Gerner; Nicholas A Gherardin; Kamran Ghoreschi; Lara Gibellini; Florent Ginhoux; Keisuke Goda; Dale I Godfrey; Christoph Goettlinger; Jose M González-Navajas; Carl S Goodyear; Andrea Gori; Jane L Grogan; Daryl Grummitt; Andreas Grützkau; Claudia Haftmann; Jonas Hahn; Hamida Hammad; Günter Hämmerling; Leo Hansmann; Goran Hansson; Christopher M Harpur; Susanne Hartmann; Andrea Hauser; Anja E Hauser; David L Haviland; David Hedley; Daniela C Hernández; Guadalupe Herrera; Martin Herrmann; Christoph Hess; Thomas Höfer; Petra Hoffmann; Kristin Hogquist; Tristan Holland; Thomas Höllt; Rikard Holmdahl; Pleun Hombrink; Jessica P Houston; Bimba F Hoyer; Bo Huang; Fang-Ping Huang; Johanna E Huber; Jochen Huehn; Michael Hundemer; Christopher A Hunter; William Y K Hwang; Anna Iannone; Florian Ingelfinger; Sabine M Ivison; Hans-Martin Jäck; Peter K Jani; Beatriz Jávega; Stipan Jonjic; 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Alicia Martínez-Romero; Glòria Martrus; Ivana Marventano; Wlodzimierz Maslinski; Giuseppe Matarese; Anna Vittoria Mattioli; Christian Maueröder; Alessio Mazzoni; James McCluskey; Mairi McGrath; Helen M McGuire; Iain B McInnes; Henrik E Mei; Fritz Melchers; Susanne Melzer; Dirk Mielenz; Stephen D Miller; Kingston H G Mills; Hans Minderman; Jenny Mjösberg; Jonni Moore; Barry Moran; Lorenzo Moretta; Tim R Mosmann; Susann Müller; Gabriele Multhoff; Luis Enrique Muñoz; Christian Münz; Toshinori Nakayama; Milena Nasi; Katrin Neumann; Lai Guan Ng; Antonia Niedobitek; Sussan Nourshargh; Gabriel Núñez; José-Enrique O'Connor; Aaron Ochel; Anna Oja; Diana Ordonez; Alberto Orfao; Eva Orlowski-Oliver; Wenjun Ouyang; Annette Oxenius; Raghavendra Palankar; Isabel Panse; Kovit Pattanapanyasat; Malte Paulsen; Dinko Pavlinic; Livius Penter; Pärt Peterson; Christian Peth; Jordi Petriz; Federica Piancone; Winfried F Pickl; Silvia Piconese; Marcello Pinti; A Graham Pockley; Malgorzata Justyna Podolska; Zhiyong Poon; Katharina Pracht; Immo Prinz; Carlo E M Pucillo; Sally A Quataert; Linda Quatrini; Kylie M Quinn; Helena Radbruch; Tim R D J Radstake; Susann Rahmig; Hans-Peter Rahn; Bartek Rajwa; Gevitha Ravichandran; Yotam Raz; Jonathan A Rebhahn; Diether Recktenwald; Dorothea Reimer; Caetano Reis e Sousa; Ester B M Remmerswaal; Lisa Richter; Laura G Rico; Andy Riddell; Aja M Rieger; J Paul Robinson; Chiara Romagnani; Anna Rubartelli; Jürgen Ruland; Armin Saalmüller; Yvan Saeys; Takashi Saito; Shimon Sakaguchi; Francisco Sala-de-Oyanguren; Yvonne Samstag; Sharon Sanderson; Inga Sandrock; Angela Santoni; Ramon Bellmàs Sanz; Marina Saresella; Catherine Sautes-Fridman; Birgit Sawitzki; Linda Schadt; Alexander Scheffold; Hans U Scherer; Matthias Schiemann; Frank A Schildberg; Esther Schimisky; Andreas Schlitzer; Josephine Schlosser; Stephan Schmid; Steffen Schmitt; Kilian Schober; Daniel Schraivogel; Wolfgang Schuh; Thomas Schüler; Reiner Schulte; Axel Ronald Schulz; Sebastian R Schulz; Cristiano Scottá; Daniel Scott-Algara; David P Sester; T Vincent Shankey; Bruno Silva-Santos; Anna Katharina Simon; Katarzyna M Sitnik; Silvano Sozzani; Daniel E Speiser; Josef Spidlen; Anders Stahlberg; Alan M Stall; Natalie Stanley; Regina Stark; Christina Stehle; Tobit Steinmetz; Hannes Stockinger; Yousuke Takahama; Kiyoshi Takeda; Leonard Tan; Attila Tárnok; Gisa Tiegs; Gergely Toldi; Julia Tornack; Elisabetta Traggiai; Mohamed Trebak; Timothy I M Tree; Joe Trotter; John Trowsdale; Maria Tsoumakidou; Henning Ulrich; Sophia Urbanczyk; Willem van de Veen; Maries van den Broek; Edwin van der Pol; Sofie Van Gassen; Gert Van Isterdael; René A W van Lier; Marc Veldhoen; Salvador Vento-Asturias; Paulo Vieira; David Voehringer; Hans-Dieter Volk; Anouk von Borstel; Konrad von Volkmann; Ari Waisman; Rachael V Walker; Paul K Wallace; Sa A Wang; Xin M Wang; Michael D Ward; Kirsten A Ward-Hartstonge; Klaus Warnatz; Gary Warnes; Sarah Warth; Claudia Waskow; James V Watson; Carsten Watzl; Leonie Wegener; Thomas Weisenburger; Annika Wiedemann; Jürgen Wienands; Anneke Wilharm; Robert John Wilkinson; Gerald Willimsky; James B Wing; Rieke Winkelmann; Thomas H Winkler; Oliver F Wirz; Alicia Wong; Peter Wurst; Jennie H M Yang; Juhao Yang; Maria Yazdanbakhsh; Liping Yu; Alice Yue; Hanlin Zhang; Yi Zhao; Susanne Maria Ziegler; Christina Zielinski; Jakob Zimmermann; Arturo Zychlinsky Journal: Eur J Immunol Date: 2019-10 Impact factor: 6.688