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Crossroad of ovarian cancer organoid culture: Single cell suspension and mechanically sheared fragment

Lanyang Li^{¹^,^§}, Jiping Liu^{¹^,^§}, Qi Cao^{²^,^§}, Yuqing Zhao^², Yanghua Shi^¹, Chen Wang^¹, Jian Zhang^¹, Mingjie Rong^¹, Xiang Tao^², Wei Deng^{³^,⁴}(), Chunhui Cai^¹(), Xinxin Han^{¹^,⁵}()

1Shanghai Lisheng Biotech, Shanghai 200092, China

2Obstetrics and Gynecology Hospital of Fudan University, Shanghai, China

3LongHua Hospital, Shanghai University of Traditional Chinese Medicine, Shanghai 200032, China

4Department of Oncology, Shanghai Ninth People’s Hospital, Shanghai Jiao Tong University School of Medicine, Shanghai 200125, China

5Organ Regeneration X Lab, LiSheng East China Institute of Biotechnology, Peking University, Nantong 226299, China

^§ Lanyang Li, Jiping Liu, and Qi Cao contributed equally to this work.

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Highlights

• Rapid and efficient generation of uniformly-sized organoids from single cell suspensions

• Consistent gene expression in two high-fidelity ovarian cancer organoid models

• Uniformly-sized ovarian cancer organoids show consistent, reliable drug responses for personalized chemotherapy resistance tests

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This study explores the cultivation of uniformly-sized ovarian cancer organoids from single-cell suspensions, addressing challenges in high-throughput drug screening and providing insights into personalized treatment strategies against chemotherapy resistance.

Abstract

Ovarian cancer, a common gynecologic tumor, is associated with a high mortality, due to challenges in early detection within the reproductive system. According to our previous research, cultivating patient-specific organoids from mechanically sheared tissues can be utilized for drug response evaluation but has limitations for high-throughput screening efficiency due to their inconsistent size. In this research, we focused on organoids developed from single-cell suspensions to address the critical requirement for uniformity in organoid size. By the day 3 of culture, single-cell suspensions rapidly and spontaneously aggregated into spherical structures with a more consistent size. Notably, the organoids of sample OVA-37 were ten times larger after 8 days of culture. Transcriptomic analysis was used to compare the two organoid culture techniques, demonstrating that the variations between different organoid culture methods were minimal, with higher variability observed among patients. Gene set enrichment analysis (GSEA) revealed only minor discrepancies in specific pathways, such as TGF-β and tight junctions. Furthermore, treatment with carboplatin in a 96-well plate setup resulted in reproducible drug responses, as evidenced by coefficients of variation lower than 40%. This finding suggests that single-cell suspension-cultured organoids can be employed for reproducible high-throughput drug screening. This approach holds potential for personalized drug screening in ovarian cancer and may contribute to the development of novel therapeutic strategies.

Keywords

ovarian cancer organoid drug sensitive personalized medicine

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Cell Organoid

DOI: 10.26599/CO.2024.9410005

Cite this article:

Li L, Liu J, Cao Q, et al. Crossroad of ovarian cancer organoid culture: Single cell suspension and mechanically sheared fragment. Cell Organoid, 2024, https://doi.org/10.26599/CO.2024.9410005