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Publication
Isolation, culture, and differentiation of Blastema cells from the regenerating caudal fin of zebrafish
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Advisor(s)
Abstract(s)
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 undi erentiated cells,
called blastema, proliferate beneath the wound-epidermis and di erentiate 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 di erentiation.
Upon treatment with all-trans retinoic acid, zebrafish blastema cells di erentiated 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 di erentiation. Furthermore, this protocol is simple, rapid, and cost-e cient, and can be virtually
applied to the development of any fish blastema cell culture.
Description
Keywords
Primary cell culture Zebrafish Blastema Cell diferentiation Fin regeneration
