Willemze R, Lurvink E, Bakker W, Journée H
Eur J Cancer Clin Oncol. 1985 Jan;21(1):127-34. doi: 10.1016/0277-5379(85)90209-3.
A modification of the in vivo tumor clonogenic assay using plasma clot diffusion chambers has been described which allows improved study of the cytological characteristics of the colonies cultured. Seventy-five percent of the tumors derived from malignant ascites could be cultured successfully (more than 30 small and large colonies per diffusion chamber). The cloning efficiency ranged from 0.01 to 10%. The addition of 2-mercaptoethanol, horse serum and insulin to the diffusion chambers did not affect colony formation, whereas the effect of cell-free malignant ascites added to the diffusion chambers was unpredictable. Colony growth was comparable when fresh or cryopreserved tumor cells were cultured. A linear relationship between the number of tumor cells inoculated and the number of colonies cultured was apparent in the range 10(2)-10(4) cells. Colony formation was stimulated by pre-irradiation (8 Gy) of the host animal and by weekly transplantation of the diffusion chambers in new mice. Intravenously administered doxorubicin penetrated the plasma clot and caused inhibition of colony formation in two experiments with melanoma cells.
已描述了一种使用血浆凝块扩散小室对体内肿瘤克隆形成试验的改进方法，该方法能更好地研究培养菌落的细胞学特征。源自恶性腹水的肿瘤中有75%能够成功培养（每个扩散小室有30多个大小菌落）。克隆效率在0.01%至10%之间。向扩散小室中添加2-巯基乙醇、马血清和胰岛素不影响菌落形成，而添加无细胞恶性腹水对扩散小室的影响则不可预测。培养新鲜或冻存的肿瘤细胞时，菌落生长情况相当。在接种的肿瘤细胞数量为10(2)-10(4)个细胞的范围内，接种的肿瘤细胞数量与培养的菌落数量之间呈明显的线性关系。宿主动物预先照射（8 Gy）以及每周将扩散小室移植到新的小鼠体内可刺激菌落形成。在两项针对黑色素瘤细胞的实验中，静脉注射阿霉素可穿透血浆凝块并导致菌落形成受到抑制。
