Further Development of an Equine Cell Line that can be Propagated over 100 Times.
Abstract: Cell lines originating from horses are necessary for isolation and propagation of equine herpesviruses (EHV). Although we established an equine-derived cell line, FHK-Tcl3, propagation ceased after fewer than 40 passages. In this study, FHK-Tcl3 cell propagation continued beyond 40 passages, achieving over 100 passages. FHK-Tcl3 cells were then cloned by limiting dilution at the 100th passage. Cloned cells were termed FHK-Tcl3.1. FHK-Tcl3.1 cells grew well and were propagated every 3 to 4 days by splitting 1:5. In addition, EHV-1, -2 and -4 showed a clear cytopathic effect (CPE) in FHK-Tcl3.1 cells, and this CPE was very similar to those seen in parental FHK-Tcl3 and primary fetal horse kidney cells. FHK-Tcl3.1 cells continue to propagate and the current passage record is over 100 times after cloning. Therefore, this cell appears to have been immortalized. FHK-Tcl3.1 cells have potential for growth and diagnosis of various equine viruses, including equine herpesviruses.
Publication Date: 2009-07-15 PubMed ID: 24833964PubMed Central: PMC4013958DOI: 10.1294/jes.20.11Google Scholar: Lookup
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- Journal Article
Summary
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The research article discusses the significant development of a horse cellular line (FHK-Tcl3.1) which can be propagated beyond 100 times and can be employed for the isolation and cultivation of equine herpes viruses.
Objective of the Research
- The primary aim of this research was to improve the propagation of the FHK-Tcl3 cell line beyond its earlier limit of 40 passages.
- Another essential goal was to create a clone of the FHK-Tcl3 cell line at the 100th passage in order to boost its long-term propagation.
Procedure and Results
- The researchers managed to enhance the propagation of the FHK-Tcl3 cell line beyond 40 passages, eventually reaching more than 100 passages.
- Once the 100th passage was achieved, the FHK-Tcl3 cells were cloned by performing a limited dilution. The resulting cloned cells were labelled FHK-Tcl3.1.
- The FHK-Tcl3.1 cell line showed considerable growth and could be propagated every 3 to 4 days with a 1:5 splitting.
Implications
- A clear cytopathic effect (CPE) was noted when the FHK-Tcl3.1 cells interacted with different types of equine herpes viruses, EHV -1, -2 and -4. The observed CPE was very similar to the effect seen in parental FHK-Tcl3 and primary foetal horse kidney cells, providing evidence for the viability and utility of the new cell line.
- The cloned FHK-Tcl3.1 cells have been found to propagate beyond 100 times, providing clear indications of being immortalized.
- Due to its potential for growth and the record for sustained propagation, the FHK-Tcl3.1 cell line can significantly contribute to the diagnosis and study of various equine viral diseases, notably equine herpesviruses.
Cite This Article
APA
Andoh K, Kai K, Matsumura T, Maeda K.
(2009).
Further Development of an Equine Cell Line that can be Propagated over 100 Times.
J Equine Sci, 20(2), 11-14.
https://doi.org/10.1294/jes.20.11 Publication
Researcher Affiliations
- Laboratory of Veterinary Microbiology, Faculty of Agriculture, Yamaguchi University, 1677-1 Yoshida, Yamaguchi 753-8515, Japan.
- Laboratory of Veterinary Microbiology, Faculty of Agriculture, Yamaguchi University, 1677-1 Yoshida, Yamaguchi 753-8515, Japan.
- Molecular Biology Division, Epizootic Research Center, Equine Research Institute, Japan Racing Association, Shimotsuke, Tochigi 329-0412, Japan.
- Laboratory of Veterinary Microbiology, Faculty of Agriculture, Yamaguchi University, 1677-1 Yoshida, Yamaguchi 753-8515, Japan.
References
This article includes 11 references
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Citations
This article has been cited 3 times.- Furusawa Y, Yamada S, Itai S, Nakamura T, Yanaka M, Sano M, Harada H, Fukui M, Kaneko MK, Kato Y. PMab-219: A monoclonal antibody for the immunohistochemical analysis of horse podoplanin. Biochem Biophys Rep 2019 Jul;18:100616.
- Mahmoud HY, Andoh K, Hattori S, Terada Y, Noguchi K, Shimoda H, Maeda K. Characterization of glycoproteins in equine herpesvirus-1. J Vet Med Sci 2013 Oct;75(10):1317-21.
- Liu D, Zhao X, Wang X. The Genomic Characterization of Equid Alphaherpesviruses: Structure, Function, and Genetic Similarity. Vet Sci 2025 Mar 3;12(3).
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