Establishment and cryopreservation of a cell line derived from caudal fin of endangered catfish Clarias dussumieri Valenciennes, 1840

We describe a new cell line, Clarias dussumieri fin (ClDuF), from the caudal fin of C. dussumieri using the explant technique followed by cryopreservation. The cryopreserved CiDuF cells were validated for quality and other characteristics. They showed typical epithelial morphology in vitro and epith...

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Bibliographic Details
Published in:Journal of fish biology 2020-03, Vol.96 (3), p.722-730
Main Authors: Swaminathan, T. Raja, Dharmaratnam, Arathi, Raja, S. Arun, Raj, N. Sundar, Lal, Kuldeep K
Format: Article
Language:English
Subjects:
Biotechnology
Catfish
Cell culture
cell line
characteristics
Chromosomes
Clarias dussumieri
Cloning
Contamination
Cryopreservation
Cytokeratin
Cytology
Diploids
Epithelial cells
Fins
Freezing
Freshwater fishes
Genes
Genetic resources
Mitochondria
Morphology
rRNA 16S
Somatic cells
Viability
Viruses
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Summary:We describe a new cell line, Clarias dussumieri fin (ClDuF), from the caudal fin of C. dussumieri using the explant technique followed by cryopreservation. The cryopreserved CiDuF cells were validated for quality and other characteristics. They showed typical epithelial morphology in vitro and epithelial cells outgrew their fibroblast cells after the fifth passage. ClDuF cells had a characteristic sigmoid curve with population doubling in 24 h. Immunotyping of the ClDuF cells against cytokeratin suggested the epithelial lineage. Chromosome analysis showed normal diploid (2n = 50) numbers and the cells did not contain any contamination, including Mycoplasma and other microbes. Partial sequencing of fragments of mitochondrial 16s rRNA and COI genes of ClDuF confirmed that the cell line was initiated from C. dussumieri. Cells at the 10th and 25th passages had more than 80% and 70% viability in the culture, respectively, after 6 months of storage at LN2. These ClDuF cells were morphologically identical to the cells before freezing and the genetic resource of C. dussumieri was preserved. The species‐specific cells can serve as a valuable source for virus isolation, conservation and cloning of somatic cells.
ISSN:0022-1112
1095-8649
DOI:10.1111/jfb.14265