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Technical Note

Isolation, Culture, and Differentiation of Blastema Cells from the Regenerating Caudal Fin of Zebrafish

1
Centre for Ocean Research, Sathyabama Institute of Science and Technology, Jeppiaar Nagar, Rajiv Gandhi Salai, Chennai, Tamil Nadu 600119, India
2
Centre of Marine Sciences (CCMAR), University of Algarve, Campus de Gambelas, 8005-139 Faro, Portugal
3
Department of Biomedical Sciences and Medicine (DCBM) and Algarve Biomedical Center (ABC), University of Algarve, Campus de Gambelas, 8005-139 Faro, Portugal
*
Author to whom correspondence should be addressed.
Received: 3 July 2019 / Revised: 6 January 2020 / Accepted: 19 January 2020 / Published: 30 January 2020
The caudal fin of teleost fish has become an excellent system for investigating the mechanisms of epimorphic regeneration. Upon amputation of the caudal fin, a mass of undifferentiated cells, called blastema, proliferate beneath the wound-epidermis and differentiate into various cell types to faithfully restore the missing fin structures. Here we describe a protocol that can be used to isolate and culture blastema cells from zebrafish. Primary cultures were initiated from 36 h post-amputation (hpa) blastema and optimal cell growth was achieved using L-15 medium supplemented with 5% fetal bovine serum in plates either coated with fibronectin or uncoated. After seeding, zebrafish blastema cells formed a uniform culture and exhibited polygonal shapes with prominent nucleus, while various cell types were also observed after few days in culture indicating cell differentiation. Upon treatment with all-trans retinoic acid, zebrafish blastema cells differentiated into neuron-like and oligodendritic-like cells. Immunocytochemistry data also revealed the presence of mesenchymal and neuronal cells. The availability of blastema cell cultures could contribute to a better understanding of epimorphic regeneration by providing a mean to investigate the mechanisms underlying blastema cell differentiation. Furthermore, this protocol is simple, rapid, and cost-efficient, and can be virtually applied to the development of any fish blastema cell culture. View Full-Text
Keywords: primary cell culture; blastema; zebrafish; fin regeneration; cell differentiation primary cell culture; blastema; zebrafish; fin regeneration; cell differentiation
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MDPI and ACS Style

Vijayakumar, P.; Cancela, M.L.; Laizé, V. Isolation, Culture, and Differentiation of Blastema Cells from the Regenerating Caudal Fin of Zebrafish. Fishes 2020, 5, 6. https://doi.org/10.3390/fishes5010006

AMA Style

Vijayakumar P, Cancela ML, Laizé V. Isolation, Culture, and Differentiation of Blastema Cells from the Regenerating Caudal Fin of Zebrafish. Fishes. 2020; 5(1):6. https://doi.org/10.3390/fishes5010006

Chicago/Turabian Style

Vijayakumar, Parameswaran, M. L. Cancela, and Vincent Laizé. 2020. "Isolation, Culture, and Differentiation of Blastema Cells from the Regenerating Caudal Fin of Zebrafish" Fishes 5, no. 1: 6. https://doi.org/10.3390/fishes5010006

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