Author Correction: Organoid culture media formulated with growth factors of defined cellular activity

Urbischek, Manuela; Rannikmae, Helena; Foets, Thomas; Ravn, Katharina; Hyvönen, Marko

doi:10.1038/s41598-020-68552-8

Cited by 2 publications

(2 citation statements)

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“…Organoid models hold great potential for modeling human disease and providing a highly reliable tool to investigate new therapeutic approaches. Despite improvements in defining the optimal growth conditions for PDOs (34,35), there is still space for optimization regarding tissue processing that might help to increase the success rate of CRC PDO generation. In normal conditions, the human colon is covered by a mucus layer that consists of an inner gel-like layer, and a loose outer layer.…”

Section: Discussionmentioning

confidence: 99%

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

et al. 2022

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The use of patient-derived organoids (PDO) as a valuable alternative to in vivo models significantly increased over the last years in cancer research. The ability of PDOs to genetically resemble tumor heterogeneity makes them a powerful tool for personalized drug screening. Despite the extensive optimization of protocols for the generation of PDOs from colorectal tissue, there is still a lack of standardization of tissue handling prior to processing, leading to microbial contamination of the organoid culture. Here, using a cohort of 16 patients diagnosed with colorectal carcinoma (CRC), we aimed to test the efficacy of phosphate-buffered saline (PBS), penicillin/streptomycin (P/S), and Primocin, alone or in combination, in preventing organoid cultures contamination when used in washing steps prior to tissue processing. Each CRC tissue was divided into 5 tissue pieces, and treated with each different washing solution, or none. After the washing steps, all samples were processed for organoid generation following the same standard protocol. We detected contamination in 62.5% of the non-washed samples, while the use of PBS or P/S-containing PBS reduced the contamination rate to 50% and 25%, respectively. Notably, none of the organoid cultures washed with PBS/Primocin-containing solution were contaminated. Interestingly, addition of P/S to the washing solution reduced the percentage of living cells compared to Primocin. Taken together, our results demonstrate that, prior to tissue processing, adding Primocin to the tissue washing solution is able to eliminate the risk of microbial contamination in PDO cultures, and that the use of P/S negatively impacts organoids growth. We believe that our easy-to-apply protocol might help increase the success rate of organoid generation from CRC patients.

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Section: Discussionmentioning

confidence: 99%

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

et al. 2022

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“…Lgr5 + ISCs reside at the crypt bottom where they are permanently exposed to locally secreted growthpromoting factors. Therefore, the addition of the niche factors EGF (14), Noggin (15) and R-Spondin 1 (15)(16)(17) to the culture medium promotes the proliferation of the ISC and transit-amplifying compartment cells to produce immature spheroids (12). This transitional stage is followed by the formation of mature "mini-guts" with clearly defined crypt-villus structures (12).…”

Section: Intestinal Organoids: History Of Origins and Current Fields Of Applicationmentioning

confidence: 99%

Intestinal organoids in coculture: redefining the boundaries of gut mucosa ex vivo modeling

Hentschel

Seufferlein

Armacki

2021

American Journal of Physiology-Gastrointestinal and Liver Physi

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All-time preservation of an intact mucosal barrier is crucial to ensuring intestinal homeostasis and, hence, the organism's overall health maintenance. This complex process relies on an equilibrated signaling system between the intestinal epithelium and numerous cell populations inhabiting the gut mucosa. Any perturbations of this delicate crosstalk, particularly regarding the immune cell compartment and microbiota, may sustainably debilitate the intestinal barrier function. As a final joint event, a critical rise in epithelial permeability facilitates the exposure of submucosal immunity to microbial antigens, resulting in uncontrolled inflammation, collateral tissue destruction and dysbiosis. Organoid-derived intestinal co-culture models have established themselves as convenient tools to re-enact such pathophysiological events, explore interactions between selected cell populations and assess their roles with a central focus on intestinal barrier recovery and stabilization.

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Author Correction: Organoid culture media formulated with growth factors of defined cellular activity

Abstract: An amendment to this paper has been published and can be accessed via a link at the top of the paper.

Cited by 2 publications

References 0 publications

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

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

Intestinal organoids in coculture: redefining the boundaries of gut mucosa ex vivo modeling

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