Establishment of a pluripotent embryonic cell line from sea perch (Lateolabrax japonicus) embryos

Chen, Songlin; Sha, Zhenxia; Ye, Han-Qing

doi:10.1016/s0044-8486(02)00570-7

Cited by 89 publications

(65 citation statements)

References 24 publications

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“…Sea perch early embryonic cell culture (LJES1) was established previously in our laboratory (Chen et al, 2003a). Routine culture of the LJES1 cells was performed as described (Chen et al, 2003a).…”

Section: Methodsmentioning

confidence: 99%

“…To develop ES cell lines and gene targeting technique in fish, extensive attempts have been made in model fish such as zebrafish (Danio rerio; Collodi et al, 1992;Sun et al, 1995;Fan et al, 2004a) and medaka (Oryzias latipes; Wakamatsu et al, 1994;Hong et al, 1996). In commercial fish species, ES-like cell lines have also been described in the gilthead seabream (Sparus aurata; Bejar et al, 2002), sea perch (L. japonicus; Chen et al, 2003a), red sea bream (Chrysophrys major; Chen et al, 2003b), and turbot (Scophthalmus maximus; Holen and Hamre, 2003). The medaka ES cell lines MES1 (Hong et al, 1998b) and Sa BE-1c from seabream (Bejar et al, 2002) were shown to retain a diploid karyotype and the ability to form viable chimeras.…”

Section: Introductionmentioning

confidence: 99%

“…Pluripotent embryonic cell cultures have been developed and differentiation ability in vitro of the normal cell culture LJES1 was demonstrated (Chen et al, 2003a). However, no data are available on the development of GFP-expressing cell strain, its in vitro differentiation ability, and chimera generation.…”

Section: Introductionmentioning

confidence: 99%

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Pluripotency and Chimera Competence of an Embryonic Stem Cell Line from the Sea Perch (Lateolabrax japonicus)

Chen

Sha

et al. 2006

Mar Biotechnol

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A stable GFP-expressing (GFP + LJES1) cell strain was developed from the LJES1 cells obtained from sea perch (Lateolabrax japonicus,) embryos. GFP + LJES1 cells were induced in vitro by RA to differentiate into a variety of cell types and also had the ability to form embryoid body-like structures in suspension culture. To determine the differentiation potential of LJES1 cells in vivo, GFP + LJES1 cells were transplanted into sea perch and zebrafish embryos at midblastula stage. Twenty out of 478 transplanted sea perch embryos contained GFP-expressing LJES1 cells 24 h after microinjection. Fifteen chimera embryos developed into fry. In these chimeras, the GFP + LJES1 cells contributed to a variety of tissues including the head and trunk. In zebrafish, 221 embryos were microinjected with GFP + LJES1 cells and 22 chimera embryos and fries expressing GFP were obtained. Donor GFP + LJES1 cells contributed to various tissues in head and trunk of zebrafish embryos and hatched fry.

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Section: Methodsmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

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Pluripotency and Chimera Competence of an Embryonic Stem Cell Line from the Sea Perch (Lateolabrax japonicus)

Chen

Sha

et al. 2006

Mar Biotechnol

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“…Blastocytes, which remain undifferentiated, are easier to handle than cells from younger embryos and are competent for generating germ line chimera offspring (Lin et al, 1992;Wakamatsu et al, 1993). To obtain blastocytes, eggs are dechorionized by digestion with hatching enzyme in medaka , with pronase in zebrafish (Sun et al, 1995b), or mechanically with fine forceps ( Figure 2B) in gilthead seabream (Béjar et al, 1999), red seabream (Chen et al, 2003a), and sea perch (Chen et al, 2003b). After washing and removal of the yolk, blastocytes are seeded either on a layer of feeder cells or in gelatinecoated culture wells ( Figures 1B and 2C).…”

Section: Es Cell Technology In Fish Step By Stepmentioning

confidence: 99%

Fish ES Cells and Applications to Biotechnology

et al. 2006

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ES cells provide a promising tool for the generation of transgenic animals with site-directed mutations. When ES cells colonize germ cells in chimeras, transgenic animals with modified phenotypes are generated and used either for functional genomics studies or for improving productivity in commercial settings. Although the ES cell approach has been limited to mice, there is strong interest for developing the technology in fish. We describe the step-by-step procedure for developing ES cells in fish. Key aspects include avoiding cell differentiation, specific in vitro traits of pluripotency, and, most importantly, testing for production of chimeric animals as the main evidence of pluripotency. The entire process focuses on two model species, zebrafish and medaka, in which most work has been done. The achievements attained in these species, as well as their applicability to other commercial fish, are discussed. Because of the difficulties relating to germ line competence, mostly of long-term fish ES cells, alternative cell-based approaches such as primordial germ cells and nuclear transfer need to be considered. Although progress to date has been slow, there are promising achievements in homologous recombination and alternative avenues yet to be explored that can bring ES technology in fish to fruition.

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“…Fertile nuclear transplants have also been obtained from nuclei of medaka blastulae (Wakamatsu et al, 2001) and of zebrafish fibroblast cultures (Lee et al, 2002). Much progress has been made towards the development of ES cells in medaka (Wakamatsu et al, 1994;Hong et al, , 1998a, zebrafish (Ma et al, 2001) as well as other fish species including the seabream (Bejar et al, 2002), red seabream (Chen et al, 2003a) and sea perch (Chen et al, 2003b). In particular, one medaka ES cell line, MES1, has proven to retain a diploid karyotype and the ability to form chimeric adults (Hong et al, , 1998a(Hong et al, , b, c, 2000(Hong et al, , 2003.…”

Section: Introductionmentioning

confidence: 99%

Retention of the Developmental Pluripotency in Medaka Embryonic Stem Cells after Gene Transfer and Long-term Drug Selection for Gene Targeting in Fish

et al. 2004

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Embryonic stem (ES) cells provide a unique tool for introducing random or targeted genetic alterations, because it is possible that the desired, but extremely rare recombinant genotypes can be screened by drug selection. ES cell-mediated transgenesis has so far been limited to the mouse. In the fish medaka (Oryzias latipes) several ES cell lines have been made available. Here we report the optimized conditions for gene transfer and drug selection in the medaka ES cell line MES1 as a prelude for gene targeting in fish. MES1 cells gave rise to a moderate to high transfection efficiency by the calcium phosphate co-precipitation (5%), commercial reagents Fugene (11%), GeneJuice (21%) and electroporation (>30%). Transient gene transfer and CAT reporter assay revealed that several enhancers/promoters and their combinations including CMV, RSV and ST (the SV40 virus early gene enhancer linked to the thymidine kinase promoter) were suitable regulatory sequences to drive transgene expression in the MES1 cells. We show that neo, hyg or pac conferred resistance to G418, hygromycin or puromycin for positive selection, while the HSV-tk generated sensitivity to ganciclovir for negative selection. The positive-negative selection procedure that is widely used for gene targeting in mouse ES cells was found to be effective also in MES1 cells. Importantly, we demonstrate that MES1 cells after gene transfer and long-term drug selection retained the developmental pluripotency, as they were able to undergo induced differentiation in vitro and to contribute to various tissues and organs during chimeric embryogenesis.

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Establishment of a pluripotent embryonic cell line from sea perch (Lateolabrax japonicus) embryos

Cited by 89 publications

References 24 publications

Pluripotency and Chimera Competence of an Embryonic Stem Cell Line from the Sea Perch (Lateolabrax japonicus)

Pluripotency and Chimera Competence of an Embryonic Stem Cell Line from the Sea Perch (Lateolabrax japonicus)

Fish ES Cells and Applications to Biotechnology

Retention of the Developmental Pluripotency in Medaka Embryonic Stem Cells after Gene Transfer and Long-term Drug Selection for Gene Targeting in Fish

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