Sacasg and sacalg: new GFP-labelled camel skin and lung fibroblast cell lines
| Autor: | Abdullah Alenazi, Ziyad Alhamdan, Mohammad N. Alkhrayef, Faisal Alagrafi, Abdullah Alawad, Othman Alhazzaa, Mohamed Hammad, Sultan N. Alharbi |
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| Rok vydání: | 2016 |
| Předmět: |
endocrine system
Pathology medicine.medical_specialty 040301 veterinary sciences Cell growth fungi 04 agricultural and veterinary sciences Transfection Biology Molecular biology In vitro Green fluorescent protein 0403 veterinary science medicine.anatomical_structure Cell culture Lipofectamine Gene expression medicine Animal Science and Zoology Fibroblast |
| Zdroj: | Journal of Camel Practice and Research. 23:73 |
| ISSN: | 2277-8934 0971-6777 |
| DOI: | 10.5958/2277-8934.2016.00010.2 |
| Popis: | Camelids skin is claimed to respond differently under different physiological and pathological condition, however, there is no in vitro model with a reporter to investigate this claim. So generating camel fibroblast cell lines that expresses green fluorescent protein (GFP) would be important as a tool to monitor camel cell growth, migration and other processes. Hence, in this investigation, we created a GFP expressing fibroblast cell lines derived from a primary skin and lung fibroblast cell lines of the Arabian camel with a stable expression of GFP which was transfected using the pCAG-EGFP plasmid with CMV enhancer via Lipofectamine 2000 reagent. Stably transfected clones were selected by puromycin screening. The results revealed that camel fibroblasts can be efficiently transduced in vitro using pCAG-CMV-based vectors and that these vectors direct long-term transgene expression without evident toxicity, pathogenesis or alteration of native fibroblast morphology. Immunofluorescence staining showed no differences in the expression of fibroblast biomarkers between the transfected and non-transfected cell lines. The viability of thawed cells remained above 85% after cryopreservation in liquid nitrogen. The gene expression of the fibroblast markers was not different in the transfected cell lines. Taken together, we have established and fully characterised GFP expressing-fibroblast cell lines of Arabian camel. Based on our assays, we conclude that transfection of GFP into the Arabian camel skin and lung fibroblasts did not change their observed properties. The GFP-labelled cell lines may represent a new tool for convenient monitoring of live primary camel fibroblasts. |
| Databáze: | OpenAIRE |
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