Y. Nanki, A. Hirasawa, H. Nomura, A. Okubo, Manabu Itoh, T. Akahane, T. Chiyoda, F. Kataoka, E. Tominaga, D. Aoki
{"title":"摘要A61:腹水来源和组织来源的卵巢癌细胞原代3D培养用于个性化医疗","authors":"Y. Nanki, A. Hirasawa, H. Nomura, A. Okubo, Manabu Itoh, T. Akahane, T. Chiyoda, F. Kataoka, E. Tominaga, D. Aoki","doi":"10.1158/1557-3265.OVCA17-A61","DOIUrl":null,"url":null,"abstract":"Objective: Ovarian cancer is a poor-prognosis gynecologic disease with over 220,000 diagnoses each year worldwide and a 5-year survival rate less than 40%. In this study, we present a method for isolation and characterization of ovarian cancer cells from patient-derived ascites and tissue samples by using three-dimensional (3D) cell culture. Materials and Methods: Ascites and tissue samples were obtained from primary ovarian, peritoneal, and fallopian tube cancer patients intraoperatively. Samples were isolated and cultured within 6 hours after collection. After isolation, cells were resuspended in Matrigel and were placed at the center of each well. Optimized medium is added to each well. NanoCulture Plate LH96 (Low-Binding, MH pattern, 96-wells, Medical and Biological Laboratories Co., Ltd., Japan) were used for 3D cell culture. Plates were incubated at 37°C. The cultures were examined for metabolically active cells by ATP assay and medium changed on days 0, 1, 4, 7, 10, and 14. Results: We successfully obtained ascites-derived primary cell cultures within 1-7 days from 100% (3/31) of the ascites samples and 62% (5/8) from tissue-derived primary cell cultures. Spheroids-like structures were formed in 30% (1/3) of ascites samples and 50% (4/8) of tissue samples. The tumorigenicity and invasiveness of the cells were demonstrated using new 3D spheroid model cultured in vitro by NanoCulture Plate LH96. Conclusion: Primary ascites and tissue culture of ovarian cancer cells can successfully be cultured by 3D cell culture plates. We may apply this method for drug sensitivity testing; therefore, 3D cell culture has a potential to evaluate the sensitivity of candidate chemotherapeutic drug for individual patients. Citation Format: Yoshiko Nanki, Akira Hirasawa, Hiroyuki Nomura, Aki Okubo, Manabu Itoh, Tomoko Akahane, Tatsuyuki Chiyoda, Fumio Kataoka, Eiichiro Tominaga, Daisuke Aoki. Ascites-derived and tissue-derived ovarian cancer cell primary 3D cultures aimed for personalized medicine. [abstract]. In: Proceedings of the AACR Conference: Addressing Critical Questions in Ovarian Cancer Research and Treatment; Oct 1-4, 2017; Pittsburgh, PA. Philadelphia (PA): AACR; Clin Cancer Res 2018;24(15_Suppl):Abstract nr A61.","PeriodicalId":18646,"journal":{"name":"Metabolic Changes in Ovarian Cancer","volume":"295 1","pages":""},"PeriodicalIF":0.0000,"publicationDate":"2018-08-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":"1","resultStr":"{\"title\":\"Abstract A61: Ascites-derived and tissue-derived ovarian cancer cell primary 3D cultures aimed for personalized medicine\",\"authors\":\"Y. Nanki, A. Hirasawa, H. Nomura, A. Okubo, Manabu Itoh, T. Akahane, T. Chiyoda, F. Kataoka, E. Tominaga, D. Aoki\",\"doi\":\"10.1158/1557-3265.OVCA17-A61\",\"DOIUrl\":null,\"url\":null,\"abstract\":\"Objective: Ovarian cancer is a poor-prognosis gynecologic disease with over 220,000 diagnoses each year worldwide and a 5-year survival rate less than 40%. In this study, we present a method for isolation and characterization of ovarian cancer cells from patient-derived ascites and tissue samples by using three-dimensional (3D) cell culture. Materials and Methods: Ascites and tissue samples were obtained from primary ovarian, peritoneal, and fallopian tube cancer patients intraoperatively. Samples were isolated and cultured within 6 hours after collection. After isolation, cells were resuspended in Matrigel and were placed at the center of each well. Optimized medium is added to each well. NanoCulture Plate LH96 (Low-Binding, MH pattern, 96-wells, Medical and Biological Laboratories Co., Ltd., Japan) were used for 3D cell culture. Plates were incubated at 37°C. The cultures were examined for metabolically active cells by ATP assay and medium changed on days 0, 1, 4, 7, 10, and 14. Results: We successfully obtained ascites-derived primary cell cultures within 1-7 days from 100% (3/31) of the ascites samples and 62% (5/8) from tissue-derived primary cell cultures. Spheroids-like structures were formed in 30% (1/3) of ascites samples and 50% (4/8) of tissue samples. The tumorigenicity and invasiveness of the cells were demonstrated using new 3D spheroid model cultured in vitro by NanoCulture Plate LH96. Conclusion: Primary ascites and tissue culture of ovarian cancer cells can successfully be cultured by 3D cell culture plates. We may apply this method for drug sensitivity testing; therefore, 3D cell culture has a potential to evaluate the sensitivity of candidate chemotherapeutic drug for individual patients. Citation Format: Yoshiko Nanki, Akira Hirasawa, Hiroyuki Nomura, Aki Okubo, Manabu Itoh, Tomoko Akahane, Tatsuyuki Chiyoda, Fumio Kataoka, Eiichiro Tominaga, Daisuke Aoki. Ascites-derived and tissue-derived ovarian cancer cell primary 3D cultures aimed for personalized medicine. [abstract]. In: Proceedings of the AACR Conference: Addressing Critical Questions in Ovarian Cancer Research and Treatment; Oct 1-4, 2017; Pittsburgh, PA. Philadelphia (PA): AACR; Clin Cancer Res 2018;24(15_Suppl):Abstract nr A61.\",\"PeriodicalId\":18646,\"journal\":{\"name\":\"Metabolic Changes in Ovarian Cancer\",\"volume\":\"295 1\",\"pages\":\"\"},\"PeriodicalIF\":0.0000,\"publicationDate\":\"2018-08-01\",\"publicationTypes\":\"Journal Article\",\"fieldsOfStudy\":null,\"isOpenAccess\":false,\"openAccessPdf\":\"\",\"citationCount\":\"1\",\"resultStr\":null,\"platform\":\"Semanticscholar\",\"paperid\":null,\"PeriodicalName\":\"Metabolic Changes in Ovarian Cancer\",\"FirstCategoryId\":\"1085\",\"ListUrlMain\":\"https://doi.org/10.1158/1557-3265.OVCA17-A61\",\"RegionNum\":0,\"RegionCategory\":null,\"ArticlePicture\":[],\"TitleCN\":null,\"AbstractTextCN\":null,\"PMCID\":null,\"EPubDate\":\"\",\"PubModel\":\"\",\"JCR\":\"\",\"JCRName\":\"\",\"Score\":null,\"Total\":0}","platform":"Semanticscholar","paperid":null,"PeriodicalName":"Metabolic Changes in Ovarian Cancer","FirstCategoryId":"1085","ListUrlMain":"https://doi.org/10.1158/1557-3265.OVCA17-A61","RegionNum":0,"RegionCategory":null,"ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":null,"EPubDate":"","PubModel":"","JCR":"","JCRName":"","Score":null,"Total":0}
引用次数: 1
摘要
目的:卵巢癌是一种预后不良的妇科疾病,全球每年有超过22万例诊断,5年生存率不到40%。在这项研究中,我们提出了一种通过三维(3D)细胞培养从患者来源的腹水和组织样本中分离和表征卵巢癌细胞的方法。材料与方法:术中获取原发性卵巢、腹膜和输卵管癌患者的腹水和组织样本。样品在采集后6小时内分离培养。分离后,细胞在Matrigel中重悬,并置于每孔的中心。每口井都添加了优化的介质。采用纳米培养板LH96 (Low-Binding, MH pattern, 96-wells, Medical and Biological Laboratories Co., Ltd, Japan)进行三维细胞培养。37℃孵育。通过ATP法检测培养物的代谢活性细胞,并在第0、1、4、7、10和14天更换培养基。结果:我们在1-7天内成功获得100%(3/31)的腹水来源的原代细胞培养,62%(5/8)的组织来源的原代细胞培养。30%(1/3)的腹水样本和50%(4/8)的组织样本形成球状结构。利用纳米培养板LH96体外培养新的三维球体模型,证实了细胞的致瘤性和侵袭性。结论:三维细胞培养板可成功培养卵巢癌细胞原代腹水和组织培养。本方法可用于药敏试验;因此,3D细胞培养有可能评估候选化疗药物对个体患者的敏感性。引文格式:南木良子、平泽明、野村博之、大久保明、伊藤真昭、赤韩知子、千代田达之、片冈文雄、富永英一郎、青木大辅。腹水来源和组织来源的卵巢癌细胞原代3D培养旨在个性化医疗。[摘要]。AACR会议论文集:解决卵巢癌研究和治疗中的关键问题;2017年10月1-4日;宾夕法尼亚州匹兹堡。费城(PA): AACR;临床肿瘤杂志,2018;24(15 -增刊):摘要nr A61。
Abstract A61: Ascites-derived and tissue-derived ovarian cancer cell primary 3D cultures aimed for personalized medicine
Objective: Ovarian cancer is a poor-prognosis gynecologic disease with over 220,000 diagnoses each year worldwide and a 5-year survival rate less than 40%. In this study, we present a method for isolation and characterization of ovarian cancer cells from patient-derived ascites and tissue samples by using three-dimensional (3D) cell culture. Materials and Methods: Ascites and tissue samples were obtained from primary ovarian, peritoneal, and fallopian tube cancer patients intraoperatively. Samples were isolated and cultured within 6 hours after collection. After isolation, cells were resuspended in Matrigel and were placed at the center of each well. Optimized medium is added to each well. NanoCulture Plate LH96 (Low-Binding, MH pattern, 96-wells, Medical and Biological Laboratories Co., Ltd., Japan) were used for 3D cell culture. Plates were incubated at 37°C. The cultures were examined for metabolically active cells by ATP assay and medium changed on days 0, 1, 4, 7, 10, and 14. Results: We successfully obtained ascites-derived primary cell cultures within 1-7 days from 100% (3/31) of the ascites samples and 62% (5/8) from tissue-derived primary cell cultures. Spheroids-like structures were formed in 30% (1/3) of ascites samples and 50% (4/8) of tissue samples. The tumorigenicity and invasiveness of the cells were demonstrated using new 3D spheroid model cultured in vitro by NanoCulture Plate LH96. Conclusion: Primary ascites and tissue culture of ovarian cancer cells can successfully be cultured by 3D cell culture plates. We may apply this method for drug sensitivity testing; therefore, 3D cell culture has a potential to evaluate the sensitivity of candidate chemotherapeutic drug for individual patients. Citation Format: Yoshiko Nanki, Akira Hirasawa, Hiroyuki Nomura, Aki Okubo, Manabu Itoh, Tomoko Akahane, Tatsuyuki Chiyoda, Fumio Kataoka, Eiichiro Tominaga, Daisuke Aoki. Ascites-derived and tissue-derived ovarian cancer cell primary 3D cultures aimed for personalized medicine. [abstract]. In: Proceedings of the AACR Conference: Addressing Critical Questions in Ovarian Cancer Research and Treatment; Oct 1-4, 2017; Pittsburgh, PA. Philadelphia (PA): AACR; Clin Cancer Res 2018;24(15_Suppl):Abstract nr A61.