Growth of human small cell (oat cell) carcinoma of the lung in serum-free growth factor-supplemented medium.
Growth of human small cell (oat cell) carcinoma of the lung in serum-free growth factor-supplemented medium.
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发表时间：
1980-12
期刊：
影响因子：
11.2
通讯作者：
E. Simms;A. Gazdar;P. Abrams;J. Minna
中科院分区：
文献类型：
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作者：
E. Simms;A. Gazdar;P. Abrams;J. Minna
摘要研究了人肺小细胞癌细胞系NCI-H69在无血清培养基中复制所需生长因子。通过一系列添加实验，发现将硒（3 × 10 - 8 m）、氢化可的松（10 - 8 m）、胰岛素（5 μg/ml）、转铁蛋白（100 μg/ml）和17β-雌二醇（10 - 8 m）（SHITE）加到不含血清的Roswell Park Memorial Institute Medium 1640中，当细胞从含血清培养基中转移时，可获得最佳复制。NCI-H69细胞在补充SHITE的罗斯威尔公园纪念研究所培养基1640中连续复制超过12个月。细胞复制与补充有10%胎牛血清的培养基中大致相同的倍增时间，但它们表现出较低的饱和密度和较长的滞后期在SHITE补充的培养基相比，血清补充的培养基。当删除SHITE的单个组分时，仅在删除转铁蛋白的情况下观察到生长的立即和显著差异，而在删除其他因子的情况下观察到最小的降低。在SHITE中生长6个月后，从配方中删除胰岛素或转铁蛋白导致无血清培养基中细胞生长停止，而删除氢化可的松或雌二醇（单独或一起）则不会。各种其他激素和生长因子并没有显着增加SHITE在促进复制。此外，发现一系列激素抑制复制或饱和密度达到无血清，补充维生素E的培养基，包括表皮生长因子，促黄体生成素释放因子，甲状旁腺激素，血粉，和三肽甘氨酰-L-组氨酰-L-赖氨酸乙酸酯。测试了其他六个小细胞癌系，并且也能够在补充有SHITE的无血清培养基中复制，表明该配方对于小细胞癌的通用性。在SHITE培养基中培养4个月的NCI-H69细胞保留了它们的胺前体摄取和脱羧特性，通过光学显微镜观察到的特征性小细胞癌组织学，致密核心（神经分泌）颗粒，通过电子显微镜，和裸鼠的致瘤性。该生长公式的鉴定应允许直接从患者样品中选择肿瘤细胞，研究这些细胞系的激素产生和调节，并鉴定新的生长和治疗剂，并且它应提供一种选择与小细胞癌组织遗传学相关的正常胺处理细胞的方法。
Abstract A cell culture line of human small cell (oat cell) carcinoma of the lung (NCI-H69) was studied for growth factor requirements for replication in serum-free medium. By a series of addition experiments, the combination of selenium (3 × 10 -8 m), hydrocortisone (10 -8 m), insulin (5 µg/ml), transferrin (100 µg/ml), and 17β-estradiol (10 -8 m) (SHITE) added to Roswell Park Memorial Institute Medium 1640 without serum was found to allow optimum replication when cells were transferred from serum-containing medium. NCI-H69 cells have replicated continuously in SHITE-supplemented Roswell Park Memorial Institute Medium 1640 for periods of more than 12 months. The cells replicate with approximately the same doubling times as in medium supplemented with 10% fetal calf serum, but they exhibited a lower saturation density and longer lag phase in SHITE-supplemented medium compared to serum-supplemented medium. When individual components of SHITE were deleted, immediate and dramatic differences in growth were seen only with deletion of transferrin while minimal decreases were noted with deletion of the other factors. After 6 months of growth in SHITE, deletion of insulin or transferrin from the formula resulted in cessation of cell growth in serum-free medium while deletion of hydrocortisone or estradiol (alone or together) did not. A variety of other hormones and growth factors did not add significantly to SHITE in promoting replication. In addition, a series of hormones was found to inhibit replication or saturation density achieved in serum-free, hormone-supplemented medium including epidermal growth factor, luteinizing hormone-releasing factor, parathyroid hormone, blood meal, and the tripeptide glycyl-l-histidyl-l-lysine acetate. Six other small cell carcinoma lines were tested and also were able to replicate in SHITE-supplemented serum-free medium indicating the generality of this formula for small cell carcinoma, NCI-H69 cells cultured for 4 months in SHITE medium retained their amine precursor uptake and decarboxylation properties, characteristic small cell carcinoma histology by light microscopy, dense core (neurosecretory) granules by electron microscopy, and tumorigenicity in nude mice. The identification of this growth formula should allow selection of tumor cells directly from patient samples, study of hormone production and regulation by these cell lines, and identification of new growth and therapeutic agents, and it should provide a way of selecting for normal amine handling cells related histogenetically to small cell carcinoma.