Viana Cristiano Ribeiro, Neto Cristovam Scapulatempo, Kerr Ligia Maria, Palmero Edenir Inêz, Marques Marcia Maria Chiquitelli, Colaiacovo Tamara, de Queiroz Junior Abel Feliciano, Carvalho André Lopes, Siqueira Sheila Aparecida Coelho
Department of Pathology, Barretos Cancer Hospital (Pio XII Foundation), Barretos, Brazil.
Cell Tissue Bank. 2013 Jun;14(2):167-73. doi: 10.1007/s10561-012-9313-5. Epub 2012 May 5.
Tumor Banks were created to organize the collection, storage and distribution of biological samples from oncological patients, facilitating its use in cancer research. To ensure the quality of the samples from our bank, we implemented standard operating procedures international. In order to evaluate the influence of cold ischemia time (time between surgical removal of the specimen and the snap freezing of the sample) on the quality of the samples (evaluated by measurement integrity of their RNA), collected during 10 months two tumor samples from each donor, one with up to 30 min of cold ischemia and other with exact 45 min, totaling 100 different donors and 200 samples, 40 from each of the following organs: breast, thyroid, stomach, lung and colorectum. We extracted total RNA from the samples and with the aid of a Bioanalyser, evaluate their quality, comparing it with cold ischemia times in different organs. Among the samples up to 30 min and the samples with exact 45 min, we respectively found 63 (64.3 %) and 36 (36 %) with intact RNA, 11 (11.2 %) and 17 (17 %) partially degraded and 24 (24.5 %) and 47 (47 %) degraded (p < 0.001). Thyroid and colorectal samples were more sensitive to variations in cold ischemia time (p = 0.006 and p = 0.03, respectively). Stomach and lungs were less sensitive (p = 0.919 and p = 0.384, respectively). We concluded that the cold ischemia time up to 30 min was more efficient to maintain the integrity of RNA in most samples, and that RNA degradation varied according to the different topographies.
肿瘤库的建立是为了组织收集、储存和分发肿瘤患者的生物样本，以促进其在癌症研究中的应用。为确保我们库中样本的质量，我们实施了国际标准操作程序。为了评估冷缺血时间（从手术切除标本到样本速冻之间的时间）对样本质量（通过测量其RNA完整性来评估）的影响，在10个月内，从每个供体收集两个肿瘤样本，一个冷缺血时间长达30分钟，另一个正好45分钟，共有100个不同的供体和200个样本，分别来自以下器官：乳腺、甲状腺、胃、肺和结肠直肠，每个器官40个样本。我们从样本中提取总RNA，并借助生物分析仪评估其质量，将其与不同器官的冷缺血时间进行比较。在冷缺血时间长达30分钟的样本和正好45分钟的样本中，我们分别发现63个（64.3%）和36个（36%）样本的RNA完整，11个（11.2%）和17个（17%）样本部分降解，24个（24.5%）和47个（47%）样本降解（p<0.001）。甲状腺和结肠直肠样本对冷缺血时间的变化更敏感（分别为p = 0.006和p = 0.03）。胃和肺样本较不敏感（分别为p = 0.919和p = 0.384）。我们得出结论，在大多数样本中，长达30分钟的冷缺血时间更有利于维持RNA的完整性，并且RNA降解因不同的部位而异。
