An efficient and user‐friendly method for cytohistological analysis of organoids

Zhang, Shiwei; Chen, Wei; Lu, Xiufen; Zhang, Wen; Hu, Lei; Liu, Yuhong; Yang, Zheng; Xue, Ling; Qiao, Shan; Liang, Yan; Oliveira, Joaquím M.; Reis, Rui L.; Zhang, Changhua

doi:10.1002/term.3248

Cited by 7 publications

(4 citation statements)

References 24 publications

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“…These tiny organoids, ranging from micrometer to millimeter scale, have become a model of choice for many applications, but their small sizes are often a technical challenge for their histological analysis using classic embedding methods such as paraffin. Some improvement techniques have been proposed to obtain better histology, including the use of agarose as a pre-embedding step to concentrate the organoids 21 , 24 , but with these experiments, the organoids are fixed and cannot be reused later on for viable assays. Here we demonstrated the potential of stimulated Raman histology (SRH) on rectal cancer patients-derived organoids to provide a three-dimensional histopathology without any perturbation of the biological functionality of the living cells.…”

Section: Introductionmentioning

confidence: 99%

In vivo organoid growth monitoring by stimulated Raman histology

Sarri,

Chevrier,

Poizat

et al. 2024

npj Imaging

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Patient-derived tumor organoids have emerged as a crucial tool for assessing the efficacy of chemotherapy and conducting preclinical drug screenings. However, the conventional histological investigation of these organoids necessitates their devitalization through fixation and slicing, limiting their utility to a single-time analysis. Here, we use stimulated Raman histology (SRH) to demonstrate non-destructive, label-free virtual staining of 3D organoids, while preserving their viability and growth. This novel approach provides contrast similar to conventional staining methods, allowing for the continuous monitoring of organoids over time. Our results demonstrate that SRH transforms organoids from one-time use products into repeatable models, facilitating the efficient selection of effective drug combinations. This advancement holds promise for personalized cancer treatment, allowing for the dynamic assessment and optimization of chemotherapy treatments in patient-specific contexts.

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Section: Introductionmentioning

confidence: 99%

In vivo organoid growth monitoring by stimulated Raman histology

Sarri,

Chevrier,

Poizat

et al. 2024

npj Imaging

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“…The culture protocol for colorectal cancer organoids was based primarily on previously published work [18], and the complete formulation of culture medium can be seen in the supporting information. For tumor organoid extraction, the patientderived tumor samples were rinsed with a Tissue Washing Solution (EACIN BIO).…”

Section: Organoid and Cell Line Culturementioning

confidence: 99%

3D bioprinting platform development for high-throughput cancer organoid models construction and drug evaluation

Dai,

Chen,

Chen

et al. 2024

Biofabrication

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The evaluation of anti-tumor drugs is critical for their development and clinical guidance. Tumor organoid models are gaining increased attention due to their ability to better mimic real tumor tissues, as well as lower time and economic costs, which makes up for the shortcomings of cell lines and xenograft models. However, current tumor organoid cultures based on the Matrigel have limitations in matching with high-throughput engineering methods due to slow gelation and low mechanical strength. Here, we present a novel composite bioink for culturing colorectal cancer organoids that provides an environment close to real tissue growth conditions and exhibits excellent photocrosslinking properties for rapid gel formation. Most importantly, the tumor organoids viability in the composite bioink after printing was as high as 97%, which also kept multicellular polar structures consistent with traditional culture methods in the Matrigel. Using 3D bioprinting with this composite bioink loaded with organoids, we demonstrated the feasibility of this drug evaluation model by validating it with clinically used colorectal cancer treatment drugs. Our results suggested that the composite bioink could effectively cultivate tumor organoids using 3D bioprinting, which had the potential to replace less reliable manual operations in promoting the application of tumor organoids in drug development and clinical guidance.

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“…The frozen sections were prepared using a previously published protocol. 28 The organoid cultures' medium was discarded, and Matrigel was digested in Cell Recovery solution at 4°C for 30 min. The supernatant was removed after centrifugation.…”

Section: Real-time Quantitative Polymerase Chain Reactionmentioning

confidence: 99%

Regulation of epithelial cell differentiation by the Ubiquitous expressed transcript isoform 1 in ulcerative colitis

Peng,

Zeng,

et al. 2023

J of Gastro and Hepatol

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Background and AimMucosal healing has emerged as a desirable treatment goal for patients with ulcerative colitis (UC). Healing of mucosal wounds involves epithelial cell proliferation and differentiation, and Y‐box transcription factor ZONAB has recently been identified as the key modulator of intestinal epithelial restitution.MethodsWe studied the characteristics of UXT‐V1 expression in UC patients using immunohistochemistry and qPCR. The functional role of UXT‐V1 in the colonic epithelium was investigated using lentivirus‐mediated shRNA in vitro and ex vivo. Through endogenous Co‐immunoprecipitation and LC–MS/MS, we identified ZONAB as a UXT‐V1‐interactive protein.ResultsHerein, we report that UXT‐V1 promotes differentiation of intestinal epithelial cells by regulating the nuclear translocation of ZONAB. UXT‐V1 was upregulated in the intestinal epithelia of UC patients compared with that of healthy controls. Knocking down UXT‐V1 in NCM‐460 cells led to the enrichment of pathways associated with proliferation and differentiation. Furthermore, the absence of UXT‐V1 in cultured intestinal epithelial cells and colonic organoids inhibited differentiation to the goblet cell phenotype. Mechanistically, the loss of UXT‐V1 in the intestinal epithelial cells allowed nuclear translocation of ZONAB, wherein it regulated the transcription of differentiation‐related genes, including AML1 and KLF4.ConclusionTaken together, our study reveals a potential role of UXT‐V1 in regulating epithelial cell differentiation, proving a molecular basis for mucosal healing in UC.

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An efficient and user‐friendly method for cytohistological analysis of organoids

Cited by 7 publications

References 24 publications

In vivo organoid growth monitoring by stimulated Raman histology

In vivo organoid growth monitoring by stimulated Raman histology

3D bioprinting platform development for high-throughput cancer organoid models construction and drug evaluation

Regulation of epithelial cell differentiation by the Ubiquitous expressed transcript isoform 1 in ulcerative colitis

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