Feb 13, 2026

Public workspaceConstruction of Matrigel encapsulated PDOs (ME-PDOs)

DOI
https://dx.doi.org/10.17504/protocols.io.261ge1jnwv47/v1
  • Zhe Zhao1
  • 1CAS Key Laboratory of Nano-Bio Interface, Suzhou Institute of Nano-Tech and Nano-Bionics, Chinese Academy of Sciences, Jiangsu, 215123, China.
  • MTS
Guangli Suo
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DOI: https://dx.doi.org/10.17504/protocols.io.261ge1jnwv47/v1
Protocol Citation: Zhe Zhao 2026. Construction of Matrigel encapsulated PDOs (ME-PDOs). protocols.io https://dx.doi.org/10.17504/protocols.io.261ge1jnwv47/v1
License: This is an open access protocol distributed under the terms of the Creative Commons Attribution License,  which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited
Protocol status: Working
We use this protocol and it's working
Created: February 13, 2026
Last Modified: February 13, 2026
Protocol Integer ID: 243178
Keywords: derived organoid, construction of matrigel, clinical breast cancer, pdo, resected clinical breast cancer, matrigel
Funders Acknowledgements:
National Key Research and Development Program of China
Grant ID: 2022YFC2705005
Abstract
Establishment of patient-derived organoids (PDOs) from freshly resected clinical breast cancer specimens.
Troubleshooting
Construction of Matrigel encapsulated PDOs (ME-PDOs)
The BRC tissues were digested as described above. Briefly, the tissues were digested on a shaker at 37 °C for 1–2 hours with intermittent pipetting until the visible pieces disappeared.
Dissociated cell clusters were spun down by centrifugation at 1,200 rpm and 4 °C for 5 minutes. The resulting dissociated cell clusters or single cells were resuspended in cold Matrigel (Corning) diluted to a concentration of 50% and seeded in prewarmed 24-well plate using drops of 25 µL each.
The drops were solidified in a 5% CO₂ incubator at 37 °C for 30 minutes, followed by addition of 1.5 mL organoid culture medium (bioGenous) to each well, which was refreshed every 2 to 3 days for around 2 weeks totally. Then the ME-PDOs were digested into single ME-PDO for further test.
Protocol references
1. Sachs, N. et al. A Living Biobank of Breast Cancer Organoids Captures Disease Heterogeneity. Cell 172, 373-386 e310 (2018).
2. Broutier, L. et al. Human primary liver cancer-derived organoid cultures for disease modeling and drug screening. Nat Med 23, 1424-1435 (2017).