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Related Experiment Video

Updated: Oct 5, 2025

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Standardizing Patient-Derived Organoid Generation Workflow to Avoid Microbial Contamination From Colorectal Cancer

Mattia Marinucci1, Caner Ercan1,2, Stephanie Taha-Mehlitz1,3

  • 1Visceral Surgery and Precision Medicine Research Laboratory, Department of Biomedicine, University of Basel, Basel, Switzerland.

Frontiers in Oncology
|January 27, 2022
PubMed
Summary
This summary is machine-generated.

Primocin effectively prevents microbial contamination in patient-derived organoid (PDO) cultures from colorectal cancer (CRC) tissues. This washing step enhances organoid generation success without negatively impacting cell viability, unlike penicillin/streptomycin.

Keywords:
Primocinantibioticscolorectal cancermicrobial contamination controlpatient-derived organoids

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Area of Science:

  • Cancer Research
  • Cell Biology
  • Microbiology

Background:

  • Patient-derived organoids (PDOs) are valuable for personalized cancer drug screening due to their genetic similarity to tumors.
  • Standardization of tissue handling before processing is lacking, leading to microbial contamination in colorectal cancer (CRC) organoid cultures.
  • Existing protocols need optimization to prevent contamination and ensure successful organoid generation.

Purpose of the Study:

  • To evaluate the efficacy of different washing solutions in preventing microbial contamination during colorectal cancer organoid culture.
  • To compare the impact of phosphate-buffered saline (PBS), penicillin/streptomycin (P/S), and Primocin on organoid contamination rates and cell viability.

Main Methods:

  • Colorectal carcinoma (CRC) tissues from 16 patients were divided into five pieces each.
  • Tissue pieces were treated with PBS, P/S, Primocin, combinations, or no solution before organoid processing.
  • Organoid cultures were generated using a standardized protocol, and contamination rates were assessed.

Main Results:

  • Unwashed samples showed a 62.5% contamination rate; PBS and P/S-containing PBS reduced it to 50% and 25%, respectively.
  • Washing with PBS/Primocin completely eliminated contamination in 100% of organoid cultures.
  • Penicillin/streptomycin (P/S) significantly reduced the percentage of viable cells compared to Primocin.

Conclusions:

  • Primocin in the washing solution is highly effective in preventing microbial contamination in patient-derived organoid (PDO) cultures.
  • The addition of P/S to washing solutions negatively impacts organoid cell viability.
  • This optimized washing protocol can improve the success rate of generating PDOs from colorectal cancer patients.