Basic Research
Establishment and preliminary application of organoids in ovarian cancer
Zhang Lin, Meng Huangyang, Zhang Yashuang, Miao Huixian, Yuan Lin, Zhou Shulin, Jiang Yi, Wan Yicong, Cheng Wenjun
Published 2023-02-25
Cite as Chin J Obstet Gynecol, 2023, 58(2): 112-120. DOI: 10.3760/cma.j.cn112141-20221103-00677
Abstract
ObjectiveTo explore the establishment and application of ovarian cancer organoids.
MethodsFresh ovarian tumor tissues, obtaining from patients underwent surgery in the First Affiliated Hospital of Nanjing Medical University between October 2021 and March 2022, were collected, enzymatic degraded, digested, and embedded into matrigel to establish organoids. A total of 32 ovarian cancer samples were collected. Hematoxylin eosin (HE) staining and immunofluorescence (IF) procedure were used to verify the morphological structure of organoids and their expression of molecular markers. 3D cyto-live or dead assay was used to detecte the live or dead cells in organoids. Carboplatin with a concentration ranging from 5 to 80 μmol/L (5, 10, 20, 40, 80 μmol/L) was added to organoids to calculate the 50% inhibitory concentration (IC50) in different organoids.
Results(1) Organoids from a total of 32 patients were established, of which 18 cases could be passaged stably in the long term in vitro, while 14 could be passaged in the short time. The average amplification time of long-term passage in vitro was over 3 months, and the longest reached 9 months. (2) In HE staining, significant nuclei atypia and local micropapillary structures were observed in organoids. IF staining revealed that ovarian cancer organoids expressed molecular markers similar to primary tumor tissues, such as Pan cytokeratin (Pan-CK), p53, paired box gene 8 (PAX8), and Wilms tumor gene 1 (WT1). (3) In 3D cyto-live or dead assay, a large number of apoptotic cells were observed inside and around the organoids after added carboplatin. The sensitivity to carboplatin varied in 18 organoids could amplify in the long term, with an average IC50 of (29.5±15.8) μmol/L. Moreover, IC50 values of 4 organoids derived from patients received neoadjuvant chemotherapy were much higher than the 14 organoids which did not received neoadjuvant chemotherapy [(48.7±11.3) μmol/L vs (24.0±12.1) μmol/L; t=3.429, P=0.022].
ConclusionsOrganoids recapitulate ovarian cancers in vitro and could be stably passaged. Organoids derived from patients received neoadjuvant chemotherapy have higher resistance to carboplatin.
Key words:
Ovarian neoplasms; Organoids; Chemoradiotherapy, adjuvant; Drug resistance, neoplasm; Carboplatin; Drug screening assays, antitumor
Contributor Information
Zhang Lin
Department of Gynecology, the First Affiliated Hospital of Nanjing Medical University, Nanjing 210029, China
Meng Huangyang
Department of Gynecology, the First Affiliated Hospital of Nanjing Medical University, Nanjing 210029, China
Zhang Yashuang
Department of Gynecology, the First Affiliated Hospital of Nanjing Medical University, Nanjing 210029, China
Miao Huixian
Department of Gynecology, the First Affiliated Hospital of Nanjing Medical University, Nanjing 210029, China
Yuan Lin
Department of Gynecology, the First Affiliated Hospital of Nanjing Medical University, Nanjing 210029, China
Zhou Shulin
Department of Gynecology, the First Affiliated Hospital of Nanjing Medical University, Nanjing 210029, China
Jiang Yi
Department of Gynecology, the First Affiliated Hospital of Nanjing Medical University, Nanjing 210029, China
Wan Yicong
Department of Gynecology, the First Affiliated Hospital of Nanjing Medical University, Nanjing 210029, China
Cheng Wenjun
Department of Gynecology, the First Affiliated Hospital of Nanjing Medical University, Nanjing 210029, China
