PUBLICATION
Establishment and characterization of a new fibroblast-like cell line from the skin of a vertebrate model, zebrafish (Danio rerio)
- Authors
- Sathiyanarayanan, A., Yashwanth, B.S., Pinto, N., Thakuria, D., Chaudhari, A., Gireesh Babu, P., Goswami, M.
- ID
- ZDB-PUB-221028-2
- Date
- 2022
- Source
- Molecular biology reports 50(1): 19-29 (Journal)
- Registered Authors
- Chaudhari, Aparna
- Keywords
- Bacterial extracellular products, Cell line, Danio rerio, Mycoplasma, Skin, Transfection
- MeSH Terms
-
- Aeromonas hydrophila
- Animals
- Cell Line
- Humans
- RNA, Ribosomal, 16S/genetics
- RNA, Ribosomal, 16S/metabolism
- Skin*
- Zebrafish*/genetics
- Zebrafish*/metabolism
- PubMed
- 36289143 Full text @ Mol. Biol. Rep.
Citation
Sathiyanarayanan, A., Yashwanth, B.S., Pinto, N., Thakuria, D., Chaudhari, A., Gireesh Babu, P., Goswami, M. (2022) Establishment and characterization of a new fibroblast-like cell line from the skin of a vertebrate model, zebrafish (Danio rerio). Molecular biology reports. 50(1):19-29.
Abstract
Background The available fully sequenced genome and genetic similarities compared to humans make zebrafish a prominent in vitro vertebrate model for drug discovery & screening, toxicology, and radiation biology. Zebrafish also possess well developed immune systems which is ideal for studying infectious diseases. Fish skin confers immunity by serving as a physical barrier against the invading pathogens in the aquatic habitat. Therefore in vitro models from the skin tissue of zebrafish help to study the physiology, functional genes in vitro, wound healing, and pathogenicity of microbes. Hence the study aimed to develop and characterize a skin cell line from the wild-type zebrafish Danio rerio.
Methods and results A novel cell line designated as DRS (D. rerio skin) was established and characterized from the skin tissue of wild-type zebrafish, D. rerio, by the explant technique. The cells thrived well in the Leibovitz's -15 medium supplemented with 15% FBS and routinely passaged at regular intervals. The DRS cells mainly feature fibroblast-like morphology. The culture conditions of the cells were determined by incubating the cells at varying concentrations of FBS and temperature; the optimum was 15% FBS and 28 °C, respectively. Cells were cryopreserved and revived with 70-75% viability at different passage levels. Two extracellular products from bacterial species Aeromonas hydrophila and Edwardsiella tarda were tested and found toxic to the DRS cells. Mitochondrial genes, namely COI and 16S rRNA PCR amplification and partial sequencing authenticated the species of origin of cells. The modal diploid (2n) chromosome number of the cells was 50. The cell line DRS was found to be free from mycoplasma. The cells were transfected with pMaxGFP plasmid and tested positive for green fluorescence at 24-48 h post-transfection.
Conclusion The findings from this study thus confirm the usefulness of the developed cell line in bacterial susceptibility and transgene expression studies.
Genes / Markers
Expression
Phenotype
Mutations / Transgenics
Human Disease / Model
Sequence Targeting Reagents
Fish
Orthology
Engineered Foreign Genes
Mapping