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Updated: Nov 29, 2025
Generation of Induced-pluripotent Stem Cells Using Fibroblast-like Synoviocytes Isolated from Joints of Rheumatoid Arthritis Patients
Published on: October 16, 2016
Rheumatoid Arthritis: Applicability of Ready-to-Use Human Cartilaginous Cells for Screening of Compounds with
Larissa T Volova1, Evgeniy I Pugachev1, Victoria V Rossinskaya1
1Institute of Experimental Medicine and Biotechnology, Samara State Medical University, Chapaevskaya St., 89, 443099 Samara, Russia.
Abstract:
In the context of modern drug discovery, there is an obvious advantage to designing phenotypic bioassays based on human disease-relevant cells that express disease-relevant markers. The specific aim of the study was to develop a convenient and reliable method for screening compounds with Tumor Necrosis Factor-alpha (TNF-α) inhibitory activity. This assay was developed using cryopreserved ready-to-use cartilage-derived cells isolated from juvenile donors diagnosed with polydactyly. It has been demonstrated that all donor (10 donors) cells were able to respond to TNF-α treatment by increased secretion of pro-inflammatory cytokine IL-6 into subcultural medium. Inhibition of TNF-α using commercially available TNF-α inhibitor etanercept resulted in a dose-dependent decrease in IL-6 production which was measured by Enzyme-Linked Immunosorbent Assay (ELISA). TNF-α dependent IL-6 production was detected in the cells after both their prolonged cultivation in vitro (≥20 passages) and cryopreservation. This phenotypic bioassay based on ready-to-use primary human cells was developed for detection of novel TNF-α inhibitory compounds and profiling of biosimilar drugs.
Insights
A new bioassay uses human cartilage cells to screen for Tumor Necrosis Factor-alpha (TNF-α) inhibitors. This reliable method effectively measures compounds that reduce inflammation, aiding drug discovery for TNF-α related conditions.
Area of Science:
- Biochemistry
- Cell Biology
- Pharmacology
Background:
- Phenotypic bioassays are crucial for drug discovery, particularly those using human disease-relevant cells and markers.
- Tumor Necrosis Factor-alpha (TNF-α) is a key pro-inflammatory cytokine implicated in various diseases.
- Developing reliable screening methods for TNF-α inhibitors is essential for therapeutic advancements.
Purpose of the Study:
- To develop a convenient and reliable phenotypic bioassay for screening compounds with TNF-α inhibitory activity.
- To utilize cryopreserved, ready-to-use primary human cartilage-derived cells for this assay.
- To validate the assay's performance using a known TNF-α inhibitor.
Main Methods:
- Primary human cartilage-derived cells from juvenile donors were used.
- Cells were treated with TNF-α, and the resulting IL-6 secretion was measured.
- Enzyme-Linked Immunosorbent Assay (ELISA) was employed to quantify IL-6 levels.
- The effect of etanercept, a TNF-α inhibitor, on IL-6 production was assessed.
Main Results:
- All tested donor cells (n=10) responded to TNF-α by increasing IL-6 secretion.
- Etanercept demonstrated a dose-dependent inhibition of TNF-α-induced IL-6 production.
- The assay was effective with cells after prolonged in vitro cultivation (≥20 passages) and cryopreservation.
Conclusions:
- A robust phenotypic bioassay using primary human cartilage cells was successfully developed.
- This assay is suitable for screening novel TNF-α inhibitory compounds.
- The method is valuable for profiling biosimilar drugs targeting TNF-α pathways.
