An organoid culture assay (OCA) for determining the drug sensitivity of human tumors

Abstract

A model for testing chemotherapeutic agents in vitro is described. It is based on an organoid culture method which allows human carcinomas to grow in vitro and to maintain many typical in vivo properties, including 3-dimensional architecture, growth of multiple cell types, expression of morphological differentiation and formation of histotypical structures. The preservation of drug sensitivity and resistance under the conditions of our organoid culture assay (OCA) was demonstrated by investigating 3 strains of a human hypopharynx carcinoma which differed by different sensitivity to cisplatin in in vivo conditions. These differences were retained in vitro and the modified neutral-red (NR) assay was especially suitable for revealing drug-induced cytotoxic damage in OCA. On the basis of our findings, the following approach is proposed for the in vitro testing of cytostatic drugs before they are administered to patients. (1) Removal of carcinomas from patients; (2) dense cell suspensions of these carcinomas to be dropped on membrane filters at the air-medium interface, resulting in growth of solid nodules of organized carcinoma tissues; (3) addition of cytostatic drugs to the growth medium for 2 or 3 days; (4) detachment and bisection of the culture nodules; (5) determination of viable cells by NR uptake and total cell mass by the sulforhodamin B (SRB) assay; (6) determination of quotient NR:SRB absorbance, related as percentage to control value: this indicates the fraction of viable cells and gives a measure of the cytotoxic injury caused by the applied cytotoxic drug. Thus, the OCA seems to be suitable for defining the patterns of drug sensitivity and resistance of individual human carcinomas in vitro within a few days.

Feasibility of Primary Tumor Culture Models and Preclinical Prediction Assays for Head and Neck Cancer: A Narrative Review

“…Until now, only one group described an “organoid culture assay” of HNSCC [ 86 , 87 ]. Although the authors did not use fresh primary tumor, they aimed for 3D in vitro tumor growth allowing to form organized and differentiated structures such as those existing in the organism.…”

“…After 2 weeks of spheroid formation, a prediction assay warrants another 2 weeks of incubation. Within the first weeks a decreasing proportion of epithelial cells was seen [ 80 ] and a degeneration of histological characteristics [ 86 ].…”

Preclinical models of head and neck squamous cell carcinoma for a basic understanding of cancer biology and its translation into efficient therapies

“…They showed that these organoid cultures maintained the critical properties of the in vivo state, such as the 3D architecture, the growth of heterogeneous cell types from an individual carcinoma and the morphological differentiation under relatively simple experimental conditions [7]. In a subsequent study, the same group demonstrated that these organoid cultures can be used for drug testing, and that the response data obtained thereof were concordant with patients' response to therapy [21]. The authors were the first to propose organoid cultures as personalized in vitro drug testing platform, allowing the prediction of individual chemosensitivity of carcinomas within few days [21].…”

“…In a subsequent study, the same group demonstrated that these organoid cultures can be used for drug testing, and that the response data obtained thereof were concordant with patients' response to therapy [21]. The authors were the first to propose organoid cultures as personalized in vitro drug testing platform, allowing the prediction of individual chemosensitivity of carcinomas within few days [21].…”

Review Article

“…In light of this, National Cancer Institute (NCI) announced abolishment of using NCI-60 cancer cell lines for future drug screening tests [12]. 3D organoid culture was first developed in 1950s [13,14], but did not gain attention in drug screening until recent years [15,16]. The mimicry of 3D environment in organoid cultures is its major virtue, and proves it to be more closely representative of its primary tumor.…”