In vivo bromodeoxyuridine labelling in human tumour xenografts.
In vivo bromodeoxyuridine labelling in human tumour xenografts.
复制标题
人体肿瘤异种移植物中的体内溴脱氧尿苷标记。
DOI：
10.1080/09553008914552051
复制
发表时间：
1989
影响因子：
2.6
通讯作者：
Steel,GG
中科院分区：
文献类型：
--
作者：
Parkins,CS;Begg,AC;Bush,C;Price,P;Imrie,PR;Ormerod,MG;Steel,GG
2.材料和方法培养来源于两种人宫颈肿瘤（命名为HX 156和HX 171，Kelland等，1987）的连续细胞系，并在体外与10μM BUdR在37 ℃下混合孵育30分钟。然后从该培养基中取出细胞，重悬于新鲜培养基中，立即和每隔2小时取样。将细胞固定在70%乙醇中，并保留约105个细胞用于FCM分析。对于体内实验，将人乳腺（HX 99）、膀胱（BT 14）和宫颈肿瘤（HX 155 + 156）的异种移植物在研究所设施中饲养的免疫缺陷雄性小鼠中连续传代。在肿瘤大小为约8 mm GMD（几何平均直径）时，用100 mg/kg 5-溴-2-脱氧尿苷（Sigma）腹膜内注射小鼠，并且在注射后的适当时间，将小鼠杀死并移除肿瘤，或者用戊巴比妥钠（100 mg/kg）麻醉并移除两个楔形活检组织中的第一个。对于第二个程序，将小鼠在温暖的床上保持麻醉，直到第二次活检时间过去。将肿瘤块切成5 mm 3的立方体，并立即固定在70%的冷乙醇中。根据Schutte等人的方法制备肿瘤细胞核。（1987），并用大鼠抗溴脱氧尿苷单克隆抗体（ICR 1，Sera Labs，Crawley Down，UK）染色，然后用荧光素化兔抗大鼠IgG t染色。联合王国，1989年4月11日至15日。
2. Materials and methods Continuous cell lines derived from two human cervix tumours (designated HX156 and HX171, Kelland et al. 1987) were cultured and incubated in vitro with 10µM BUdR with mixing at 37 C for 30 min. The cells were then removed from this medium and resuspended in fresh medium from which samples were taken immediately and at 2 hour intervals. The cells were fixed in 70 per cent ethanol and approximately 105 cells kept for the FCM analysis. For in vivo experiments, xenografts of human breast (HX99), bladder (BT14) and cervical tumours (HX155+ 156), were serially passaged in immune-deficient male mice, bred in the Institute's facility. At a tumour size of about 8 mm GMD (geometric mean diameter) the mice were injected ip with 100 mg/kg 5-bromo-2-deoxyuridine (Sigma) and at appropriate times after injection the mice were either killed and the tumour removed or were anaesthetised with sodium pentobarbitone (100 mg/kg) and the first of two wedge biopsies removed. For this second procedure the mice were maintained under anaesthetic on a warm bed until the second biopsy time had elapsed. Tumour pieces were chopped into 5 mm3 cubes and fixed immediately in 70 per cent cold ethanol. Tumour nuclei were prepared according to the method of Schutte et al.(1987) and stained using a rat monoclonal antibody against bromodeoxyuridine (ICR1, Sera Labs, Crawley Down, UK) followed by fluoresceinated rabbit anti-rat IgG t Presented at the 15th LH Gray ConferenceRadiobiology of Human Cells and Tissues', Canterbury. UK, 11-15 April 1989.