Bjerkvig R, Tønnesen A, Laerum O D, Backlund E O
Department of Pathology, University of Bergen School of Medicine, Haukeland Hospital, Norway.
J Neurosurg. 1990 Mar;72(3):463-75. doi: 10.3171/jns.1990.72.3.0463.
Tumor tissue from seven human gliomas was maintained in long-term agar overlay culture as multicellular organotypic spheroids. Light microscopic and ultrastructural observation of the spheroids displayed morphological features similar to those of the original tumor tissue in vivo; in this respect they were different from spheroids obtained from permanent cell lines. The spheroids contained preserved vessels, connective tissue, and macrophages, revealing a close resemblance to the conditions in the original tumor. Flow cytometric deoxyribonucleic acid measurements of cells from the tumor spheroids and from biopsy material obtained directly from the operation revealed the same ploidy and the same amount of proliferating cells in the spheroids as in the original tumor. Fluorescence microscopy using bromodeoxyuridine (BUdR) incorporation and anti-BUdR monoclonal antibody confirmed the proliferative potential of tumor cells in the spheroids. Diameter measurements showed that the size of the spheroids from two of the tumors increased over time while in three other cases it decreased. Spheroids from the remaining two tumors showed no change in size, even after 80 days in culture. These growth data and the relatively high number of proliferating cells, as measured by flow cytometry, indicate that the degree of cell proliferation and cell loss from the spheroids are closely linked, as is the case for tumors in vivo. The culture system presented provides a valuable alternative to propagation of human tumors in animals.
取自7例人类神经胶质瘤的肿瘤组织以多细胞器官样球体形式在长期琼脂覆盖培养中维持。对这些球体进行光镜和超微结构观察，显示出与体内原始肿瘤组织相似的形态特征；在这方面，它们不同于从永久细胞系获得的球体。这些球体含有保留的血管、结缔组织和巨噬细胞，与原始肿瘤的情况极为相似。对肿瘤球体细胞以及直接从手术中获取的活检材料进行流式细胞术脱氧核糖核酸测量，结果显示球体中的细胞倍性和增殖细胞数量与原始肿瘤相同。使用溴脱氧尿苷（BUdR）掺入和抗BUdR单克隆抗体的荧光显微镜检查证实了球体中肿瘤细胞的增殖潜力。直径测量表明，来自其中两个肿瘤的球体大小随时间增加，而在其他三个病例中则减小。来自其余两个肿瘤的球体即使在培养80天后大小也没有变化。这些生长数据以及通过流式细胞术测量的相对较高的增殖细胞数量表明，球体中的细胞增殖程度和细胞丢失密切相关，体内肿瘤也是如此。所呈现的培养系统为在动物体内繁殖人类肿瘤提供了一种有价值的替代方法。
