Short-term spheroid culture of primary colorectal cancer cells as an in vitro model for personalizing cancer medicine

Jeppesen, Maria; Hagel, Grith; Glenthøj, Anders; Vainer, Ben; Ibsen, Per; Harling, Henrik; Thastrup, Ole; Jørgensen, Lars Nannestad; Thastrup, Jacob

doi:10.1371/journal.pone.0183074

Cited by 73 publications

(59 citation statements)

References 55 publications

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“…Tumoroids were cultured based on the work by Jeppesen et al Tumour specimens were cut into 1‐2 mm 2 fragments using an ordinary scalpel while keeping the tissue on ice. Tumour fragments were washed with cold DPBS and dissociated partially by 1 mg/mL collagenase type II (Thermo Fisher Scientific) in DPBS at 37°C in several steps.…”

Section: Methodsmentioning

confidence: 99%

KRAS mutations in the parental tumour accelerate in vitro growth of tumoroids established from colorectal adenocarcinoma

Mousavi

Truelsen²,

Hagel³

et al. 2019

Int J Experimental Path

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The aim of the present study was to characterize a patient-derived in vitro 3D model (ie tumoroid) established from colorectal adenocarcinoma. This study investigated the growth rate of tumoroids and whether the Kirsten rat sarcoma (KRAS) mutations in the parental tumour accelerate this rate. The tumoroids were established from surgical resections of primary and metastatic colorectal adenocarcinoma from 26 patients. The in vitro growth rate of these tumoroids was monitored by automated imaging and recorded as relative growth rate. The KRAS hotspot mutations were investigated on the parental tumours by Ion Torrent ™ next-generation sequencing. The KRAS mutations were detected in 58% of the parental tumours, and a significantly higher growth rate was observed for tumoroids established from the KRAS-mutated tumours compared to wild-type tumours (P < 0.0001). The average relative growth rate (log10) on day 10 was 0.360 ± 0.180 (mean ± SD) for the KRAS-mutated group and 0.098 ± 0.135 (mean ± SD) for the KRAS wild-type group. These results showed that the presence of KRAS mutations in parental tumours is associated with an acceleration of the growth rate of tumoroids. The future perspective for such a model could be the implementation of chemoassays for personalized medicine. K E Y W O R D S 3D in vitro culture, colorectal cancer, gastrointestinal pathology, Kirsten rat sarcoma mutation, tumoroids | 15 MOUSAVI et Al.The age of the patients varied from 52 to 85 years old with an average of 70.2 years (± 8.7, SD).

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Section: Methodsmentioning

confidence: 99%

KRAS mutations in the parental tumour accelerate in vitro growth of tumoroids established from colorectal adenocarcinoma

Mousavi

Truelsen²,

Hagel³

et al. 2019

Int J Experimental Path

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“…Jeppesen et al. successfully formed spheroids from cells obtained from 15 of total 18 patients (83% success rate) with colorectal cancer in Petri dishes coated with agarose . Obtained spheroids consisted mostly of epithelial cells with up to 5% of fibroblast contamination and closely represented original tumor in terms of morphology and protein expression patterns ( Figure 5a).…”

Section: Dsrt On 3d Cell Culture Modelsmentioning

confidence: 99%

“…Authors observed distinct patterns of dose‐response in spheroids obtained from five different donors indicating maintenance of individual drug sensitivity profiles in spheroids during culturing period. Correlation of DSRT results with clinical outcome was not discussed . Hagemann et al.…”

Section: Dsrt On 3d Cell Culture Modelsmentioning

confidence: 99%

Section: Miniaturized Systems For Dsrt On 3d Cell Culture Modelsmentioning

confidence: 99%

“…Correlation of DSRT results with clinical outcome was not discussed. [98] Hagemann et al studied formation of spheroids from the PiCa cell line, which was generated from primary carcinoma specimens without enzymatic digestions of single cells. [99,100] Spheroids were grown using two methods-1) the method of hanging droplet, where cell aggregation is promoted by gravitational accumulation of cells in the water-air interface of droplets; and 2) ultra-low attachment plates (ULA), where cells are prevented from attaching to the surface, which leads to cell aggregation.…”

Section: Dsrt On 3d Cell Culture Modelsmentioning

confidence: 99%

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Precision Medicine in Oncology: In Vitro Drug Sensitivity and Resistance Test (DSRT) for Selection of Personalized Anticancer Therapy

Popova

Levkin

2020

Advanced Therapeutics

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Precision or personalized medicine aims to determine an optimal therapy for each individual patient. In oncology techniques such as next generation sequencing, mRNA‐sequencing, ChIP‐sequencing, and mass spectrometry are used to perform a full molecular profiling for each patient. However, it is not always possible to determine a suitable treatment for an individual cancer based on molecular profiling, mostly due to the high level of tumor heterogeneity. In vitro drug sensitivity and resistance test (DSRT) can be performed on cancer cells or tissues obtained from a patient with a panel of anticancer compounds in order to experimentally define sensitivity and resistance of each individual cancer. In combination with molecular profiling, DSRT can provide a fuller picture about the nature of disease, allowing for finding more appropriate therapy for each individual patient. In this progress report, studies describing in vitro DSRTs on 2D and 3D cell models based on patient‐derived cells are reviewed and challenges and future steps needed for the adaptation of these systems in clinics are discussed.

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Characterization of genetic intratumor heterogeneity in colorectal cancer and matching patient‐derived spheroid cultures

et al. 2017

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Patient‐derived in vitro cultures of colorectal cancer (CRC) may help guide treatment strategies prior to patient treatment. However, most previous studies have been performed on a single biopsy per tumor. The purpose of this study was to analyze multiple spatially distinct biopsies from CRCs and see how well intratumor heterogeneity (ITH) was recapitulated in matching patient‐derived spheroids. Three to five biopsies were collected from six CRC tumors. Each biopsy was split in two; one half was used for spheroid culturing, while the other half was used for DNA and RNA purification. For two patients, lymph node metastases were analyzed. Somatic mutations were called from whole exome sequencing data. Each tumor contained mutations shared across all biopsies and spheroids, including major CRC drivers such as APC, KRAS, and TP53. At the same time, all tumors exhibited ITH on both mutation and copy number level. The concordance between biopsies and spheroids ranged between 40 and 70% for coding mutations. For three patients, the biopsy and spheroid from matching areas clustered together, meaning that the spheroid resembled the area of origin more than the other areas. However, all biopsies and spheroids contained private mutations. Therefore, multiple cultures from spatially distinct sites of the tumor increase the insight into the genetic profile of the entire tumor. Molecular subtypes were called from RNA sequencing data. When based on transcripts from both cancer and noncancerous cells, the subtypes were largely independent of sampling site. In contrast, subtyping based on cancer cell transcripts alone was dependent on sample site and genetic ITH. In conclusion, all examined CRC tumors showed genetic ITH. Spheroid cultures partly reflected this ITH, and having multiple cultures from distinct tumor sites improved the representation of the genetic tumor subclones. This should be taken into account when establishing patient‐derived models for drug screening.

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Short-term spheroid culture of primary colorectal cancer cells as an in vitro model for personalizing cancer medicine

Cited by 73 publications

References 55 publications

KRAS mutations in the parental tumour accelerate in vitro growth of tumoroids established from colorectal adenocarcinoma

KRAS mutations in the parental tumour accelerate in vitro growth of tumoroids established from colorectal adenocarcinoma

Precision Medicine in Oncology: In Vitro Drug Sensitivity and Resistance Test (DSRT) for Selection of Personalized Anticancer Therapy

Characterization of genetic intratumor heterogeneity in colorectal cancer and matching patient‐derived spheroid cultures

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