3D Organoid modelling of hepatoblast-like and mesenchymal-like hepatocellular carcinoma cell lines

Aim: We wished to establish 3D organoid-like hepatocellular carcinoma (HCC) models from HCC cell lines.Methods: Hep3B, Huh7, HepG2, SNU398, SNU449, and SNU475 cell lines were inoculated into Matrigel and grown up to 9 days in hepatocyte specific or standard RPMI media. Spheroid formation was followed by light microscopy. Matrigel scaffolds were immobilized and embedded in paraffin, and sections were subjected to H&E and immunohistochemical staining for different hepatobiliary biomarkers. Stained material was examined under light microscopy and micro photo were taken.Results: Organoid-like structures were obtained successfully from all selected cell lines except mesenchymal-like SNU475 cells. Hep3B, Huh7, and HepG2 cell lines from hepatoblast-like sub-group formed compact 3D colonies and showed hepatocyte-like morphology and staining with different hepatocyte lineage markers as well as hepatobiliary progenitor markers. SNU398 and SNU449 cell lines from mesenchymal-like group formed irregular and loose 3D colonies that expressed vimentin homogeneously, but also several epithelial and hepatocyte lineage markers. The pattern of biomarker expression was unique for each cell line tested. Such features, not observed in tested monolayer cultures were confirmed with single-cell derived Hep3B cells.Conclusion: We described experimental conditions to obtain organoid-like structures from five different HCC cell lines representing hepatoblast-like and mesenchymal-like subgroups. These models are useful as an alternative to monolayer cultures to study phenotypic features of HCC cells. Our detailed analysis of biomarker expression in five different organoid-like structures provide convincing evidence for highly specific phenotypic features of these cell lines although they share some common or subtype-restricted features also

Considerations for the selection of tests for SARS-CoV-2 molecular diagnostics

During the course of 2020, the outbreak of Severe Acute Respiratory Syndrome Coronavirus-2 (SARS-CoV-2) spread rapidly across the world. Clinical diagnostic testing for SARS-Cov-2 infection has relied on the real-time Reverse Transcriptase Polymerase Chain Reaction and is considered the gold standard assay. Commercial vendors and laboratories quickly mobilised to develop diagnostic tests to detect the novel coronavirus, which was fundamentally important in the pandemic response. These SARS-Cov-2 assays were developed in line with the Food Drug Administration-Emergency Use Authorization guidance. Although new tests are continuously being developed, information about SARS-CoV-2 diagnostic molecular test accuracy has been limited and at times controversial. Therefore, the analytical and clinical performance of SARS-CoV-2 test kits should be carefully considered by the appropriate regulatory authorities and evaluated by independent laboratory validation. This would provide improved end-user confidence in selecting the most reliable and accurate diagnostic test. Moreover, it is unclear whether some of these rapidly developed tests have been subjected to rigorous quality control and assurance required under good manufacturing practice. Variable target gene regions selected for currently available tests, potential mutation in target gene regions, non-standardized pre-analytic phase, a lack of manufacturer independent validation data all create difficulties in selecting tests appropriate for different countries and laboratories. Here we provide information on test criteria which are important in the assessment and selection of SARS-CoV-2 molecular diagnostic tests and outline the potential issues associated with a proportion of the tests on the market

Evaluation of Cutaneous Manifestations According to the Time in Renal Transplant Recipients

Background and Design: This study is conducted to determine the prevalence and clinical characteristics of cutaneous manifestations in renal transplant patients.Materials and Methods: Hospital records of 116 renal transplant patients were retrospectively investigated. The data obtained from patients who had 6 months follow-up period were evaluated. There were 68 (58.6%) males and 48 (41.4%) females aged between 10 and 68 years (mean=36.6 years). Detailed dermatologic examination was performed. The patients were grouped according to gender (male-female), posttransplant period (1-5 years, 5-10 years, >10 years) and the drugs used (cyclosporin, tacrolimus, other than these two immunosuppressant drugs).Results: The most common cutaneous manifestations were infectious. The dermatological findings were onychomycosis (13), tinea pedis (9), acneiform disorders (15), and warts (9). The clinical evaluation after 6 months has also demonstrated the same result. Among the evaluated patients, 7% showed premalignant or malignant manifestations on clinical examination. According to the results obtained from the patient groups, it was found that gender, length of post-transplant period, and use of immunosuppressant drugs do not influence the clinical manifestations of patients.Conclusion: Dermatologic examinations and long-term follow-up should be performed in renal transplant patients

Altered cyclin D1 genotype distribution in human sporadic pituitary adenomas.

Background: The cyclin D1 gene (CCND1) is a proto-oncogene playing a critical role in the transition through the G1 ot the S phase of the cell cyle and is overexpressed in many tumors. G870A plymorphism at the exon4/intron4 splicing region of the CCND1 gene may play a role in pituitary tumorigenesis and invasiveness. The objective of this study was to examine CCND1 polymorphism i patients with different types sporadic pituitary adenomas

TAp73β Can Promote Hepatocellular Carcinoma Dedifferentiation.

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