Liang W, Ding F, Wang M X, Liu B L, Sun M
Institute of Biothermal Science, School of Medical Instruments and Food Engineering, University of Shanghai for Science and Technology, Shanghai, China.
Department of Pathology, Fudan University Shanghai Cancer Center, Shanghai, China.
Cryo Letters. 2017 Jul/Aug;38(4):321-329.
Snap freezing and RNAlater stabilization are methods that were wildly used in biospecimen depositories to preserve cancer tissues. Both methods have its own limitations. An ideal method for preservation of diseased tissues should permit the broader uses of stored tissue samples, including not just for molecular diagnostic analysis, histopathological evaluation, but also for the recovery of functional live cells and tissues as well as the regeneration of patient-derived xenograft (PDX) models for the drug screening study.
This study investigated molecular and pathological evaluation of cryopreserved colorectal cancer tissues, with an emphasis on effects of freezing method and cryoprotection. Global gene expression analysis with microarrays and histological examination of tissue samples were performed to compare tissue specimens after snap freezing, cryoprotectant permeation and subsequent cryopreservation.
Compared with the fresh tissue samples (immediately stabilized in RNAlater after collection), samples preserved by snap freezing exhibited the largest number of differentially-expressed genes. Some genes relate to neuron, drug addiction and drug binding, but the rest of differentially-expressed genes were functionally dispersive. Cryoprotectant permeation into tissue samples and subsequent cryopreservation via a rate-controlled freezing resulted in much less differentially expressed genes. Histological structures of tissue specimens were both well preserved by snap freezing and cryoproservation.
Snap freezing may not be as reliable as commonly considered. The pilot study demonstrates the feasibility of using cryopreservation to retain viable diseased tissues for multiple applications.
速冻和RNA later稳定化是生物样本库中广泛用于保存癌组织的方法。这两种方法都有其自身的局限性。一种理想的患病组织保存方法应允许对储存的组织样本进行更广泛的应用，不仅包括分子诊断分析、组织病理学评估，还包括功能性活细胞和组织的复苏以及用于药物筛选研究的患者来源异种移植（PDX）模型的再生。
本研究调查了冷冻保存的结直肠癌组织的分子和病理学评估，重点是冷冻方法和冷冻保护的效果。使用微阵列进行全局基因表达分析并对组织样本进行组织学检查，以比较速冻、冷冻保护剂渗透及随后冷冻保存后的组织标本。
与新鲜组织样本（采集后立即用RNA later稳定）相比，速冻保存的样本显示出最多的差异表达基因。一些基因与神经元、药物成瘾和药物结合有关，但其余差异表达基因在功能上是分散的。冷冻保护剂渗透到组织样本中并通过速率控制冷冻进行随后的冷冻保存，导致差异表达基因少得多。组织标本的组织结构通过速冻和冷冻保存均得到良好保存。
速冻可能不像通常认为的那样可靠。这项初步研究证明了使用冷冻保存来保留 viable 患病组织以用于多种应用的可行性。
Zotero 插件