Goike H M, Asplund A C, Pettersson E H, Liu L, Ichimura K, Collins V P
Ludwig Institute for Cancer Research, Stockholm Branch, Karolinska Institute, Stockholm and Unit of Tumour Pathology, Karolinska Hospital, Stockholm, Sweden.
Neuropathol Appl Neurobiol. 2000 Apr;26(2):172-6. doi: 10.1046/j.1365-2990.2000.026002172.x.
Human tumour xenografts maintained in nude mice are a valuable research tool. The passaging and maintenance of human tumour xenografts in immune-deficient animals are expensive and labour-intensive. This study presents a protocol that permits long-term cryopreservation of viable glioblastoma xenograft tissue pieces in liquid nitrogen. Twenty different human glioblastoma xenografts that have been successfully transplanted and repeatedly passaged in nude mice were cryopreserved to validate the method. Different passages were cryopreserved for up to 40 months. On retransplantation of the tumours, all cases except one grew successfully. In order to ensure that the individual tumours did grow (not induced mouse tumours) restriction fragment length polymorphism analysis with variable number of tandem repeats (VNTR) probes was carried out. The method permits the long-term storage of viable, retransplantable glioblastoma xenografts. It minimizes animal use for the maintenance of xenografts and permits cryo-back-up of valuable tumours, thus markedly reducing the cost and increasing the accessibility of human tumour xenografts as a research tool in biology and genetics.
维持在裸鼠体内的人肿瘤异种移植是一种有价值的研究工具。在免疫缺陷动物中传代和维持人肿瘤异种移植成本高昂且 labor-intensive。本研究提出了一种方案，可将存活的胶质母细胞瘤异种移植组织块在液氮中进行长期冷冻保存。对20种已成功移植并在裸鼠中反复传代的不同人胶质母细胞瘤异种移植进行冷冻保存以验证该方法。不同传代的样本冷冻保存长达40个月。再次移植肿瘤时，除一例之外所有病例均成功生长。为确保个体肿瘤确实生长（而非诱导小鼠肿瘤），使用可变串联重复序列（VNTR）探针进行了限制性片段长度多态性分析。该方法可长期保存存活的、可再次移植的胶质母细胞瘤异种移植。它将用于维持异种移植的动物使用减至最少，并允许对珍贵肿瘤进行冷冻备份，从而显著降低成本并增加人肿瘤异种移植作为生物学和遗传学研究工具的可及性。
