Department of Surgery, Memorial Sloan-Kettering Cancer Center, New York, NY, USA.
Ann Surg Oncol. 2010 Aug;17(8):2229-36. doi: 10.1245/s10434-010-0959-6. Epub 2010 Feb 17.
Tissue banking has become a major initiative at many oncology centers. The influence of warm ex-vivo ischemia times, storage times, and biobanking protocols on RNA integrity and subsequent microarray data is not well documented.
A prospective institutional review board-approved protocol for the banking of abdominal neoplasms was initiated at Memorial Sloan-Kettering Cancer Center in 2001. Sixty-four representative pancreas cancer specimens snap-frozen at various ex-vivo procurement times (< or =10 min, 11-30 min, 31-60 min, >1 h) and banked during three time periods (2001-2004, 2004-2006, 2006-2008) were processed. RNA integrity was determined by microcapillary electrophoresis using the RNA integrity number (RIN) algorithm and by results of laser-capture microdissection (LCM).
Overall, 42% of human pancreas cancer specimens banked under a dedicated protocol yielded RNA with a RIN of > or =7. Limited warm ex-vivo ischemia times did not negatively impact RNA quality (percentage of tissue with total RNA with RIN of > or =7 for < or =10 min, 42%; 11-30 min, 58%; 31-60 min, 33%; >60 min, 42%), and long-term storage of banked pancreas cancer biospecimens did not negatively influence RNA quality (total RNA with RIN of > or =7 banked 2001-2004, 44%; 2004-2006, 38%; 2006-2008, 50%). RNA retrieved from pancreatic cancer samples with RIN of > or =7 subject to LCM yielded RNA suitable for further downstream applications.
Fresh-frozen pancreas tissue banked within a standardized research protocol yields high-quality RNA in approximately 50% of specimens and can be used for enrichment by LCM. Quality of tissues of the biobank were not adversely impacted by limited variations of warm ischemia times or different storage periods. This study shows the challenges and investments required to initiate and maintain high-quality tissue repositories.
组织库已成为许多肿瘤中心的主要计划。在活体外缺血时间、储存时间和生物库方案对 RNA 完整性以及随后的微阵列数据的影响方面，记录并不完善。
2001 年，纪念斯隆-凯特琳癌症中心（Memorial Sloan-Kettering Cancer Center）启动了一项经机构审查委员会批准的前瞻性协议，用于储存腹部肿瘤。64 个代表性的胰腺癌标本在不同的活体外采集时间（<或=10 分钟、11-30 分钟、31-60 分钟、>1 小时）下进行冷冻，然后在三个时期（2001-2004 年、2004-2006 年、2006-2008 年）进行储存。使用 RNA 完整性编号（RIN）算法和激光捕获显微切割（LCM）的结果，通过微毛细管电泳测定 RNA 完整性。
总体而言，在专门的协议下储存的 42%的人胰腺癌标本产生的 RNA 的 RIN 值>或=7。有限的活体外缺血时间不会对 RNA 质量产生负面影响（在<或=10 分钟的时间内，组织总 RNA 的 RIN 值>或=7 的百分比为 42%；11-30 分钟为 58%；31-60 分钟为 33%；>60 分钟为 42%），长期储存的储存的胰腺癌生物标本也不会对 RNA 质量产生负面影响（2001-2004 年储存的 RNA 的 RIN 值>或=7 的比例为 44%；2004-2006 年为 38%；2006-2008 年为 50%）。用 RIN 值>或=7 进行 LCM 后从胰腺癌样本中提取的 RNA 适用于进一步的下游应用。
在标准化研究方案下储存的新鲜冷冻胰腺组织在大约 50%的标本中产生高质量的 RNA，并可通过 LCM 进行富集。生物库组织的质量不受有限的活体外缺血时间变化或不同储存期的影响。这项研究展示了启动和维持高质量组织库所需的挑战和投入。
