Immunochemical and molecular characterization of a novel cell line derived from the brain of Trachinotus blochii (Teleostei, Perciformes): A fish cell line with oligodendrocyte progenitor cell and tanycyte characteristics

Wen, Chiu-Ming; Wang, Chun-Shun; Chin, Tzu-Chuan; Cheng, Shih-Ting; Nan, Fan-Hua

doi:10.1016/j.cbpa.2010.02.003

Cited by 30 publications

(35 citation statements)

References 69 publications

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“…Brain-derived primary cultures and cell lines have been established for several other fish species including tilapia [19], another African cichlid [41], goldfish [42], rainbow trout [43]–[45], barramundi [46], sea bass [47] and pompano [48]. We observed that a fibroblast-like phenotype prevailed after primary cells were kept in culture conditions for extended periods of time and overcame crisis.…”

Section: Discussionmentioning

confidence: 91%

Derivation and Osmotolerance Characterization of Three Immortalized Tilapia (Oreochromis mossambicus) Cell Lines

et al. 2014

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Fish cell cultures are becoming more widely used models for investigating molecular mechanisms of physiological response to environmental challenge. In this study, we derived two immortalized Mozambique tilapia (Oreochromis mossambicus) cell lines from brain (OmB) and lip epithelium (OmL), and compared them to a previously immortalized bulbus arteriosus (TmB) cell line. The OmB and OmL cell lines were generated without or with Rho-associated kinase (ROCK) inhibitor/3T3 feeder layer supplementation. Although both approaches were successful, ROCK inhibitor/feeder layer supplementation was found to offer the advantages of selecting for epithelial-like cell type and decreasing time to immortalization. After immortalization (≥ passage 5), we characterized the proteomes of the newly derived cell lines (OmB and OmL) using LCMS and identified several unique cell markers for each line. Subsequently, osmotolerance for each of the three cell lines following acute exposure to elevated sodium chloride was evaluated. The acute maximum osmotolerance of these tilapia cell lines (>700 mOsm/kg) was markedly higher than that of any other known vertebrate cell line, but was significantly higher in the epithelial-like OmL cell line. To validate the physiological relevance of these tilapia cell lines, we quantified the effects of acute hyperosmotic challenge (450 mOsm/kg and 700 mOsm/kg) on the transcriptional regulation of two enzymes involved in biosynthesis of the compatible organic osmolyte, myo-inositol. Both enzymes were found to be robustly upregulated in all three tilapia cell lines. Therefore, the newly established tilapia cells lines represent valuable tools for studying molecular mechanisms involved in the osmotic stress response of euryhaline fish.

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Section: Discussionmentioning

confidence: 91%

Derivation and Osmotolerance Characterization of Three Immortalized Tilapia (Oreochromis mossambicus) Cell Lines

et al. 2014

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“…GB4C4 was proposed to be an immature astroglial cell line or multipotent neural progenitor cells similar to tanycytes (Wen et al 2009). SBP was thought to be a NSC line, expressing markers for both oligodendrocyte progenitor cells and tanycytes (Wen et al 2010). SBB-W1 was possibly a NSC line as well (Servili et al 2009). In this report, the development of a cell line from the American eel brain is described and designated eelB.…”

Section: Introductionmentioning

confidence: 96%

“…These are TB from tilapia (Wen et al 2008a), GBC4 from the orange-spotted grouper (Wen et al 2009), SBB-W1 from the European sea bass (Servili et al 2009), and SBP from snubnose pompano (Wen et al 2010). These cell lines have been examined for several marker sets, suggesting that they are slightly different.…”

Section: Introductionmentioning

confidence: 98%

Development of a cell line from the American eel brain expressing endothelial cell properties

Bloch

Walsh

et al. 2015

In Vitro Cell.Dev.Biol.-Animal

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A cell line (eelB) was developed from the outgrowth of adherent cells from brain explants of the American eel, Anguilla rostrata (Lesueur). EelB cells have been grown routinely in L-15 with 10% fetal bovine serum (FBS), undergone over 100 passages, and cryopreserved successfully. The cells from late-passage cultures (>45) were polygonal, formed capillary-like structures (CLS) on Matrigel, and stained immunocytochemically for von Willebrand factor (vWF) and for three tight junction proteins, zonula occludens-1 (ZO-1), claudin 3, and claudin 5. These results suggest that eelB is an endothelial cell line, one of the few from fish and the first from the brain. Despite this, eelB did not respond to 2,3,7,8-tetrachlorodibenzo-p-dioxin (TCDD) with the induction of CYP1A protein. The cells from early-passage cultures (<20) had more varied shapes and did not form CLS on Matrigel. Only cells from early-passage cultures formed in suspension three-dimensional aggregates that had some cells expressing alkaline phosphatase and nestin. These cells are thought to be neural stem cells and the aggregates neurospheres. The emergence of endothelial-like cells upon the continued subcultivation of cells from early-passage cultures that had neural stem cells has been described previously for mammals, but this is a first for teleosts. Remarkably, cells from all passage levels were stained strongly for senescence-associated β-galactosidase (SA β-Gal) activity.

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“…With the in vitro approach, no immunocytochemical staining was observed in primary cell cultures from cod skin (Rutberg et al 1995) and no positive bands were detected in Western blots of extracts from a goldfish caudal fin cell line (CCL-71) (Lessman et al 1993). However, cytoplasmic thread-like structures did stain in primary cell cultures from the cod brain (Rutberg et al 1995) and PC were revealed in a brain cell line, SBP, from the snubnose pompano (Wen et al 2010). Thus, the only in vitro success has been achieved with brain cells and whether cells from other organs express this tubulin modification is uncertain.…”

Section: Introductionmentioning

confidence: 63%

“…PC were found in cell lines from the brain (WE-brain5), caudal fin (WE-cfin11f and WE-cfin11e), liver (WE-liver3), retina (WErpe), spleen (WE-spleen6) and bulbus arteriosus (WEBA). The only previous observation of PC in fish cell cultures was in a snubnose pompano brain cell line, SBP, which had characteristics of oligodendrocyte progenitor cells (Wen et al 2010). …”

Section: Primary Cilia (Pc)mentioning

confidence: 97%

Demonstration of primary cilia and acetylated α-tubulin in fish endothelial, epithelial and fibroblast cell lines

Bols

2015

Fish Physiol Biochem

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Primary cilia (PC) were demonstrated for the first time in fish endothelial, epithelial and fibroblast cell lines through immunofluorescence staining with the monoclonal antibody, 6-11B-1, against acetylated α-tubulin. The study was carried out with eight recently developed cell lines from the walleye, Sander vitreus (Mitchill). These were three fibroblast-like cell lines, WE-cfin11f, WE-skin11f and WE-liver3 from, respectively, the caudal fin, skin and liver, and three epithelial-like cell lines, WE-cfin11e, WE-spleen6 and WErpe from, respectively, the caudal fin, spleen and retina. Also, endothelial-like WEBA from the bulbus arteriosus and glial-like WE-brain5 from the brain were used. Immunocytochemistry revealed strong staining for acetylated α-tubulin in mitotic spindles and midbodies for all cell lines, and in PC for all cell lines except WE-skin11f. Staining of cytoplasmic microtubules (fibrils) was absent in three cell lines, including WEBA, but present in the others, especially WE-skin11f, which might have obscured PC detection in these cells. Tubacin, an inhibitor of histone deacetylase 6, induced cytoplasmic fibrils in WEBA and the intensity of their staining in WE-cfin11f. These results suggest that the cell lines might differ in their deacetylase activities, making them useful for studying this tubulin modification in teleosts, as well as for studying PC.

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Immunochemical and molecular characterization of a novel cell line derived from the brain of Trachinotus blochii (Teleostei, Perciformes): A fish cell line with oligodendrocyte progenitor cell and tanycyte characteristics

Cited by 30 publications

References 69 publications

Derivation and Osmotolerance Characterization of Three Immortalized Tilapia (Oreochromis mossambicus) Cell Lines

Derivation and Osmotolerance Characterization of Three Immortalized Tilapia (Oreochromis mossambicus) Cell Lines

Development of a cell line from the American eel brain expressing endothelial cell properties

Demonstration of primary cilia and acetylated α-tubulin in fish endothelial, epithelial and fibroblast cell lines

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