Organoid in droplet: Production of uniform pancreatic cancer organoids from single cells
Cancer organoids have improved our understanding of recapitulating the histology, genotypes, and drug response of patient tumors for personalized medicine. However, the existing cancer organoids are typically grown in animal-derived matrices (e.g., Matrigel), which suffer from poor reproducibility a...
Saved in:
| Main Authors: | , , , , , , , , |
|---|---|
| Format: | Article |
| Language: | English |
| Published: |
Elsevier
2025-06-01
|
| Series: | Materials Today Bio |
| Subjects: | |
| Online Access: | http://www.sciencedirect.com/science/article/pii/S2590006425003254 |
| Tags: |
Add Tag
No Tags, Be the first to tag this record!
|
| _version_ | 1850187356875784192 |
|---|---|
| author | Haitao Liu Tingting Tao Zhongqiao Gan Yingying Xie Yaqing Wang Yizhao Yang Xu Zhang Xianliang Li Jianhua Qin |
| author_facet | Haitao Liu Tingting Tao Zhongqiao Gan Yingying Xie Yaqing Wang Yizhao Yang Xu Zhang Xianliang Li Jianhua Qin |
| author_sort | Haitao Liu |
| collection | DOAJ |
| description | Cancer organoids have improved our understanding of recapitulating the histology, genotypes, and drug response of patient tumors for personalized medicine. However, the existing cancer organoids are typically grown in animal-derived matrices (e.g., Matrigel), which suffer from poor reproducibility and low throughput due to uncontrollable origin of seed cells, undefined matrix, and manual manipulation. Here, we report a new strategy to massively generate uniform pancreatic cancer organoids (PCOs) in a droplet system from single cells. This system is composed of all-in-water fluids that allow to mildly encapsulate single tumor cell into isolated droplet, which subsequently proliferate and self-assemble into organoids, resembling the initial state of a tumor in the body. This high-throughput method can produce thousands of organoids in a single batch. The droplets can serve as templates for synthesizing defined microgels with proper stiffness similar to that of native tumors, facilitating functional expressions of PCOs. These organoids exhibit superior uniformity and controllability in terms of size and morphologies compared with organoids cultured in manually dropped Matrigel, due mainly to the controllable number of initiating cells and defined microgels. In addition, the established organoids maintain the key biomarkers of pancreatic tumor (e.g., KRT7, KRT19 and SOX9) and higher expression of genes associated with drug metabolism confirmed by RNA-seq and PCR analysis. Furthermore, they show distinguishing responses to four clinically used drugs in a reproducible manner in automatic pipetting workstation, indicating the feasibility of the proposed method in high-throughput drug testing. The established strategy has integrated the formation, 3D cultures, and analysis of PCOs derived from single cells in a whole system, which may provide a novel platform for advancing organoids research with standardized procedure in translational applications. |
| format | Article |
| id | doaj-art-95dfe7a008b54695abc52fc1004be975 |
| institution | OA Journals |
| issn | 2590-0064 |
| language | English |
| publishDate | 2025-06-01 |
| publisher | Elsevier |
| record_format | Article |
| series | Materials Today Bio |
| spelling | doaj-art-95dfe7a008b54695abc52fc1004be9752025-08-20T02:16:06ZengElsevierMaterials Today Bio2590-00642025-06-013210176510.1016/j.mtbio.2025.101765Organoid in droplet: Production of uniform pancreatic cancer organoids from single cellsHaitao Liu0Tingting Tao1Zhongqiao Gan2Yingying Xie3Yaqing Wang4Yizhao Yang5Xu Zhang6Xianliang Li7Jianhua Qin8Division of Biotechnology, Dalian Institute of Chemical Physics, Chinese Academy of Sciences, Dalian, 116023, ChinaDivision of Biotechnology, Dalian Institute of Chemical Physics, Chinese Academy of Sciences, Dalian, 116023, ChinaDivision of Biotechnology, Dalian Institute of Chemical Physics, Chinese Academy of Sciences, Dalian, 116023, China; University of Chinese Academy of Sciences, Beijing, 100049, ChinaDivision of Biotechnology, Dalian Institute of Chemical Physics, Chinese Academy of Sciences, Dalian, 116023, China; University of Chinese Academy of Sciences, Beijing, 100049, ChinaUniversity of Science and Technology of China, Hefei, 230026, China; Suzhou Institute for Advanced Research, University of Science and Technology of China, Suzhou, 215123, ChinaImperial College London, White City Campus, London, W12 0BZ, UKDivision of Biotechnology, Dalian Institute of Chemical Physics, Chinese Academy of Sciences, Dalian, 116023, ChinaDepartment of HBP Surgery, Beijing Chao Yang Hospital, the Capital Medical University, Beijing, 100020, China; Corresponding author.Division of Biotechnology, Dalian Institute of Chemical Physics, Chinese Academy of Sciences, Dalian, 116023, China; University of Chinese Academy of Sciences, Beijing, 100049, China; University of Science and Technology of China, Hefei, 230026, China; Suzhou Institute for Advanced Research, University of Science and Technology of China, Suzhou, 215123, China; Beijing Institute for Stem Cell and Regenerative Medicine, Beijing, 100101, China; Corresponding author. Division of Biotechnology, Dalian Institute of Chemical Physics, Chinese Academy of Sciences, Dalian, 116023, China.Cancer organoids have improved our understanding of recapitulating the histology, genotypes, and drug response of patient tumors for personalized medicine. However, the existing cancer organoids are typically grown in animal-derived matrices (e.g., Matrigel), which suffer from poor reproducibility and low throughput due to uncontrollable origin of seed cells, undefined matrix, and manual manipulation. Here, we report a new strategy to massively generate uniform pancreatic cancer organoids (PCOs) in a droplet system from single cells. This system is composed of all-in-water fluids that allow to mildly encapsulate single tumor cell into isolated droplet, which subsequently proliferate and self-assemble into organoids, resembling the initial state of a tumor in the body. This high-throughput method can produce thousands of organoids in a single batch. The droplets can serve as templates for synthesizing defined microgels with proper stiffness similar to that of native tumors, facilitating functional expressions of PCOs. These organoids exhibit superior uniformity and controllability in terms of size and morphologies compared with organoids cultured in manually dropped Matrigel, due mainly to the controllable number of initiating cells and defined microgels. In addition, the established organoids maintain the key biomarkers of pancreatic tumor (e.g., KRT7, KRT19 and SOX9) and higher expression of genes associated with drug metabolism confirmed by RNA-seq and PCR analysis. Furthermore, they show distinguishing responses to four clinically used drugs in a reproducible manner in automatic pipetting workstation, indicating the feasibility of the proposed method in high-throughput drug testing. The established strategy has integrated the formation, 3D cultures, and analysis of PCOs derived from single cells in a whole system, which may provide a novel platform for advancing organoids research with standardized procedure in translational applications.http://www.sciencedirect.com/science/article/pii/S2590006425003254Pancreatic cancerUniform organoidsDroplet microfluidicDefined microgelsDrug testing |
| spellingShingle | Haitao Liu Tingting Tao Zhongqiao Gan Yingying Xie Yaqing Wang Yizhao Yang Xu Zhang Xianliang Li Jianhua Qin Organoid in droplet: Production of uniform pancreatic cancer organoids from single cells Materials Today Bio Pancreatic cancer Uniform organoids Droplet microfluidic Defined microgels Drug testing |
| title | Organoid in droplet: Production of uniform pancreatic cancer organoids from single cells |
| title_full | Organoid in droplet: Production of uniform pancreatic cancer organoids from single cells |
| title_fullStr | Organoid in droplet: Production of uniform pancreatic cancer organoids from single cells |
| title_full_unstemmed | Organoid in droplet: Production of uniform pancreatic cancer organoids from single cells |
| title_short | Organoid in droplet: Production of uniform pancreatic cancer organoids from single cells |
| title_sort | organoid in droplet production of uniform pancreatic cancer organoids from single cells |
| topic | Pancreatic cancer Uniform organoids Droplet microfluidic Defined microgels Drug testing |
| url | http://www.sciencedirect.com/science/article/pii/S2590006425003254 |
| work_keys_str_mv | AT haitaoliu organoidindropletproductionofuniformpancreaticcancerorganoidsfromsinglecells AT tingtingtao organoidindropletproductionofuniformpancreaticcancerorganoidsfromsinglecells AT zhongqiaogan organoidindropletproductionofuniformpancreaticcancerorganoidsfromsinglecells AT yingyingxie organoidindropletproductionofuniformpancreaticcancerorganoidsfromsinglecells AT yaqingwang organoidindropletproductionofuniformpancreaticcancerorganoidsfromsinglecells AT yizhaoyang organoidindropletproductionofuniformpancreaticcancerorganoidsfromsinglecells AT xuzhang organoidindropletproductionofuniformpancreaticcancerorganoidsfromsinglecells AT xianliangli organoidindropletproductionofuniformpancreaticcancerorganoidsfromsinglecells AT jianhuaqin organoidindropletproductionofuniformpancreaticcancerorganoidsfromsinglecells |