Isolation, Culture, and Differentiation of Blastema Cells from the Regenerating Caudal Fin of Zebrafish
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 faithf...
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| Format: | Article |
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MDPI AG
2020-01-01
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| Series: | Fishes |
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| Online Access: | https://www.mdpi.com/2410-3888/5/1/6 |
| _version_ | 1818581664664649728 |
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| author | Parameswaran Vijayakumar M. Leonor Cancela Vincent Laizé |
| author_facet | Parameswaran Vijayakumar M. Leonor Cancela Vincent Laizé |
| author_sort | Parameswaran Vijayakumar |
| collection | DOAJ |
| description | 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-<i>trans</i> 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. |
| first_indexed | 2024-12-16T07:37:05Z |
| format | Article |
| id | doaj.art-1f295aa1278a4982a6e9a0c30f2c6211 |
| institution | Directory Open Access Journal |
| issn | 2410-3888 |
| language | English |
| last_indexed | 2024-12-16T07:37:05Z |
| publishDate | 2020-01-01 |
| publisher | MDPI AG |
| record_format | Article |
| series | Fishes |
| spelling | doaj.art-1f295aa1278a4982a6e9a0c30f2c62112022-12-21T22:39:11ZengMDPI AGFishes2410-38882020-01-01516010.3390/fishes5010006fishes5010006Isolation, Culture, and Differentiation of Blastema Cells from the Regenerating Caudal Fin of ZebrafishParameswaran Vijayakumar0M. Leonor Cancela1Vincent Laizé2Centre for Ocean Research, Sathyabama Institute of Science and Technology, Jeppiaar Nagar, Rajiv Gandhi Salai, Chennai, Tamil Nadu 600119, IndiaCentre of Marine Sciences (CCMAR), University of Algarve, Campus de Gambelas, 8005-139 Faro, PortugalCentre of Marine Sciences (CCMAR), University of Algarve, Campus de Gambelas, 8005-139 Faro, PortugalThe 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-<i>trans</i> 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.https://www.mdpi.com/2410-3888/5/1/6primary cell cultureblastemazebrafishfin regenerationcell differentiation |
| spellingShingle | Parameswaran Vijayakumar M. Leonor Cancela Vincent Laizé Isolation, Culture, and Differentiation of Blastema Cells from the Regenerating Caudal Fin of Zebrafish Fishes primary cell culture blastema zebrafish fin regeneration cell differentiation |
| title | Isolation, Culture, and Differentiation of Blastema Cells from the Regenerating Caudal Fin of Zebrafish |
| title_full | Isolation, Culture, and Differentiation of Blastema Cells from the Regenerating Caudal Fin of Zebrafish |
| title_fullStr | Isolation, Culture, and Differentiation of Blastema Cells from the Regenerating Caudal Fin of Zebrafish |
| title_full_unstemmed | Isolation, Culture, and Differentiation of Blastema Cells from the Regenerating Caudal Fin of Zebrafish |
| title_short | Isolation, Culture, and Differentiation of Blastema Cells from the Regenerating Caudal Fin of Zebrafish |
| title_sort | isolation culture and differentiation of blastema cells from the regenerating caudal fin of zebrafish |
| topic | primary cell culture blastema zebrafish fin regeneration cell differentiation |
| url | https://www.mdpi.com/2410-3888/5/1/6 |
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