THP-G8 cell line
cell line for IL-8 Luc assay
IL-8 Luc assay: Skin sensitization assay
IL-8 Luc assay is described in Annex C of OECD test guideline No. 442E for in vitro skin sensitization.
Establishment of cell line
A THP-1-derived IL-8 reporter cell line, THP-G8 was established by the Dept. of Dermatology, Tohoku University School of Medicine. (Takahashi T. et al. An in vitro test to screen skin sensitizers using a stable THP-1-derived IL-8 reporter cell line, THP-G8. Toxicol Sci, 124(2), 359-369, 2011) (International patent publication No. WO2012/002507A1)
General information of cell line
THP-G8, that harbors the SLO and SLR luciferase genes under the control of the IL-8 and GAPDH promoters, respectively.
Reagent for measurement
Tripluc® Luciferase assay reagent (TOYOBO Cat#MRA-301)
Controls
-4-Nitrobenzyl bromide (CAS:100-11-8, Aldrich Cat#N13054) -Lactic acid (CAS:50-21-5, Sigma Cat#L6661) -X-VIVOTM 15 (Lonza, 04-418Q): Chemically defined, serum-free hematopoietic cell medium.
Medium
-RPMI-1640 (GIBCO Cat#11875-093, 500 mL) -FBS (Biological Industries Cat#04-001-1E Lot:715004) -100 X concentrated antibiotic and antimycotic (10,000 U/mL of penicillin G,10,000 µg/mL of streptomycin and 25 µg/mL of amphotericin B in 0.85 % saline) (e.g., GIBCO Cat#15240-062) -G418 (CAS:108321-42-2, Nacalai Tesque Cat#16513-84) -Puromycin (CAS:58-58-2, InvivoGen Cat#ant-pr-1)
IL-8 Luc assay: OECD test guideline No. 442E
THP-G8 cells are utilized for the in vitro skin sensitisation test (IL-8 Luc Assay), which is approved as an internationally recognized testing protocol by the Organization for Economic Co-operation and Development (OECD). The THP-G8 cells are derived from human acute monocytic leukemia cell line (THP-1 cells) stably introducing luciferase genes, in which orange-emitting luciferase (Stable Luciferase Orange, SLO) is expressed under the control of interleukin-8 (IL-8) promoter, and red-emitting luciferase (Stable Luciferase Red, SLR) is expressed under the control of GAPDH promoter, used as an internal control reporter. To evaluate skin sensitisation, transcriptional activation of interleukin-8 (IL-8) promoter by test chemicals, seen in human dendritic cells in skin, is simply measured by luciferase reporter assay.
How to obtain cell line
Cell lines must be transfered under MTA (Material Transfer Agreement). Users must enter MTA with Tottori Bioscience Foundation through Biospecimen Laboratories, Inc. (agent) before order and obtain research license.
Order(s) for cell line should be placed after execution of MTA. Pre-payment is necessary.
Costs for initial transfer includes research license for two years.
Biospecimen Laboratories, Inc. will send cell lines and other products ordered by users. Users’ accounts will be used for shipment.
Users can use cell lines following standard protocols.