A High-Throughput Screening Model of the Tumor Microenvironment for Ovarian Cancer Cell Growth

Abstract

The tumor microenvironment plays an important role in the processes of tumor growth, metastasis and drug resistance. We have utilized a multilayered 3D primary cell culture model that reproduces the human ovarian cancer metastatic microenvironment to study the effect of the microenvironment on the pharmacological responses of cancer cells proliferation to different classes of drugs. A collection of oncology drugs was screened to identify compounds that inhibited the proliferation of ovarian cancer cells growing as monolayers or forming spheroids, on plastic and on a 3D microenvironment culture model of the omentum metastatic site, and also cells already in pre-formed spheroids. Target-based analysis of the pharmacological responses revealed that several classes of targets were more efficacious in cancer cells growing in the absence of the metastatic microenvironment, and other target classes were less efficacious in cancer cells in pre-formed spheres compared to forming spheroids cultures. These findings show that both the cellular context of the tumor microenvironment and cell adhesion mode have an essential role in cancer cell drug resistance. Therefore it is important to perform screens for new drugs using model systems that more faithfully recapitulate the tissue composition at the site of tumor growth and metastasis

Time-lapse imaging of HeLa spheroids in soft agar culture provides virtual inner proliferative activity

“…are slightly earlier than day 11.625 for spheroid in silico (3). Their generation ratios with low activity must be greater than those for spheroid in silico (3) because the growth source index decreases earlier.…”

“…Their generation ratios with low activity must be greater than those for spheroid in silico (3) because the growth source index decreases earlier. If the virtual inner proliferative activity is low up to the 13th generation, the ratios are 45.02% and 52.36% for in silico spheroids (1) and (2), respectively, which are slightly larger than the ratio of 41.17% for in silico spheroid (3). If the activity is low up to the 14th generation, both ratios are over 70%.…”

“…The ratio and distribution of high virtual inner proliferative activity in spheroids in silico (1)- (3) in Fig 8B and Fig 9 are similar to the proliferation zone ratio and histochemical profiles shown in Fig 10A. This also holds true for spheroid in silico (4) in Fig 8B, Fig 9 and Fig 10B. These results demonstrate that the virtual inner proliferative activity given to spheroids in silico on day 14 is compatible with the histochemical profiles of cross sections around the center of spheroids in vitro on the same day. Moreover, the histochemical profiles shown in Fig 10A were similar to those shown in the V-79 spheroids, and the proliferative zone ratio of 48.8% estimated from the spheroids in Fig 10A was close to the proliferative volume ratio of 0.60 of V-79 spheroids.…”

“…Cancer research models for screening have included the creation of spheroid microenvironments to test drug effects [1][2][3]. In one of the earlier studies using a spheroid-based screen, Friedrich et al [4] pointed out that using spheroid integrity and volume monitoring through phase-contrast imaging improved agent measurement when accompanied by the security of clonogenic survival.…”

From Chemotherapy to Combined Targeted Therapeutics: In Vitro and in Vivo Models to Decipher Intra-tumor Heterogeneity

“…In the study of tumor progression as well as response to treatment, selection of the most appropriate model is essential since the composition of the in vitro system can strongly influence the result of the analysis. Recently, Lal-Nag et al (2017) compared the effects of different experimental models on ovarian cancer drug sensitivity. In particular, they developed a 3D model comprising ovarian cancer cells alone or co-cultured with the omentum metastatic niche, and they demonstrated that different microenvironments strongly influence response to treatment.…”