Interordinal Chimera Formation Between Medaka and Zebrafish for Analyzing Stem Cell Differentiation

Hóng, Ní; Chen, Songlin; Ge, Ruowen; Song, Jianxing; Yi, Meisheng; Hong, Yunhan

doi:10.1089/scd.2011.0630

Cited by 31 publications

(30 citation statements)

References 45 publications

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“…The ability of diploid medaka ES cell line to form chimerism in the ZES1-injected embryos further corroborates with the earlier finding reporting the ability of this cell line to embrace the zebrafish developmental program. 29 We demonstrated that F 1 germline chimeras could be generated by the transplantation of GFP-ZES1 cells, which overexpressed GFP under the influence of the CMV promoter, into recipient zebrafish embryos. The transplanted GFP-ZES1 cells contributed to normal embryonic development of the recipient embryos, with GFP expression detected in regions such as the brain, eye, epidermis, fin, and heart.…”

Section: Cellsmentioning

confidence: 95%

“…29,30 Labeled ZES1 and MES1 cells were suspended in transplantation medium (TM; 100 mM NaCl, 5 mM KCl, 5 mM HEPES, pH 7.1). Zebrafish embryos were treated for 2 min with pronase E (1 mg/mL; Sigma) at 26°C for dechorionation.…”

Section: Embryonic Stem Cell Feeder-free Pluripotencymentioning

confidence: 99%

“…After injection, the embryos were incubated in zebrafish egg water at 23°C-28°C and regularly monitored under a fluorescent microscope. 29 Formation of germline chimera using GFP-ZES1 cell line ZES1 cells were transfected with the plasmid, pCMVHygEGFP, using FuGENE HD transfection reagent (Promega), and selected with Hygromycin B (250 lg/mL) to establish a stable ZES1 cell line (GFP-ZES1) that strongly expressed GFP under the regulation of the CMV promoter.…”

Section: Embryonic Stem Cell Feeder-free Pluripotencymentioning

confidence: 99%

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Derivation and Long-Term Culture of an Embryonic Stem Cell-Like Line from Zebrafish Blastomeres Under Feeder-Free Condition

Goh

Gan

et al. 2014

Zebrafish

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Existing zebrafish embryonic stem (ES) cell lines are derived and maintained using feeder layers. We describe here the derivation and long-term culture of an ES cell-like line derived from zebrafish blastomeres without the use of feeder cells. This line, designated as ZES1, has been maintained for more than 800 days in defined Dulbecco's modified Eagle's medium supplemented with fetal bovine serum, zebrafish embryo extract, trout serum, and human basic fibroblast growth factor. ZES1 cells possessed a morphology typical of ES cells, being round or polygonal in shape with a large nucleus and sparse cytoplasm and were mostly diploid. The cells formed individual colonies consisting of tightly packed cells that stained positively for alkaline phosphatase. ZES1 cells also formed embryoid bodies when transferred onto uncoated wells. The pluripotent nature of ZES1 cells was confirmed when they could be induced to differentiate in vitro into several cell types, through low-or high-density culture conditions. Treatment with retinoic acid also induced the differentiation of ZES1 cells into primarily neuronal cells. Using immunostaining and real-time polymerase chain reaction, we showed that Sox2, a known pluripotent marker in mammalian ES cells, was also present in ZES1 cells. Chimera experiments revealed that fluorescent-labeled ZES1 cells microinjected into zebrafish blastulas participated in the formation of all three germ layers. Using GFP-labeled ZES1 cells, chimera germline transmission was also demonstrated at the F 1 generation. In conclusion, ZES1 cells possess both in vitro and in vivo pluripotency characteristics, indicating that nonmammalian ES cells can be readily derived and maintained for a long term under feeder-free culture conditions.

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Section: Cellsmentioning

confidence: 95%

Section: Embryonic Stem Cell Feeder-free Pluripotencymentioning

confidence: 99%

Section: Embryonic Stem Cell Feeder-free Pluripotencymentioning

confidence: 99%

See 1 more Smart Citation

Derivation and Long-Term Culture of an Embryonic Stem Cell-Like Line from Zebrafish Blastomeres Under Feeder-Free Condition

Goh

Gan

et al. 2014

Zebrafish

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“…Observation and photography was carried out on Leica MZFIII stereomicroscope, Zeiss Axiovert invert and Axiovert upright microscopes with a Zeiss AxioCam M5Rc digital camera (Zeiss Corp., Germany) as described previously [44,45].…”

Section: Microscopymentioning

confidence: 99%

Medaka piwi is Essential for Primordial Germ Cell Migration

Li¹,

Hóng²,

Gui

et al. 2012

CMM

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Piwi controls the number of primordial germ cells (PGCs) via protecting maternal mRNA from decay and adult germ stem cell division in Drosophila. In mouse and zebrafish, piwi controls maintenance and differentiation of adult germ stem cell during gametogenesis. Whether piwi plays a role in PGC development of vertebrates remains unsolved. We addressed this issue by using medaka (Oryzias latipes) as a vertebrate model. Molecular cloning, sequence comparison and analyses of genomic organization and chromosome synteny led to the identification in this fish of a single piwi gene, called Opiwi. By RT-PCR analyses and in situ hybridization, the Opiwi transcript is maternally supplied and becomes restricted to PGCs and the central nervous system (CNS). Opiwi knockdown did not prevent PGC formation even in the absence of any somatic structures but did significantly reduce the number of PGCs in vivo and in vitro and affect the distribution of PGCs in developing embryos. Surprisingly, depletion of zygotic Opiwi severely and specifically affected PGC migration. We conclude that Opiwi is required not only for determining the PGC number but also for controlling PGC migration. Our results demonstrate that piwi plays a generally conserved role in germ cell development from Drosophila to vertebrate and a specific role in PGC migration.

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“…Medaka (Oryzias latipes) strain i 1 and transgenic line Vg were maintained under an artificial photoperiod of 14-h/10-h light/darkness at 26°C as described [40,45,46]. In medaka, sex maturation occurs until 3 months post hatching.…”

Section: Fishmentioning

confidence: 99%

Retinoic Acid Regulates Germ Gene Transcription In Vitro and Spermatogenesis in Testicular Organ Culture

Hóng¹,

Li²

2012

Biol Syst Open Access

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The adult testis is the male reproductive organ that produces sex hormones for sexual behaviours and sperm in a well-controlled process called spermatogenesis. Spermatogenesis involves self-renewal and differentiation of the male germ stem cells spermatogonia, culminating in the production of sperm for germline transmission. Retinoic acid (RA) is essential for fate decision and meiosis entry of mammalian germ cells. The microphthalmia-associated transcription factor (Mitf) is a newly identified transcriptional activator of germ genes in the fish medaka (Oryzias latipes). Here we report an essential role of RA on Mitf-activated germ gene expression and spermatogenesis. Medaka germ gene promoters DAZ, DND and VAS were found to contain RA-responsive elements and to exhibit little Mitf-activated transcriptional activity in response to RA treatment in both medaka ES cells and male germ stem cells. A procedure for testicular organ culture was established, which allows for test-tube spermatogenesis over a 21-day period. By the ordered testicular architecture, cellular morphology and VAS-driven GFP expression, different spermatogenic stages were identifiable on testicular cross-sections. In organ culture, RA treatment generated testes completely free of spermatogonia, and reduced VAS-driven GFP expression in spermatogonia but enhanced its expression in meiotic and post-meiotic germ cells. Therefore, RA represses Mitf-activated germ gene expression in stem cell cultures and differentially regulates germ gene expression depending upon the stages of spermatogenesis, and represses spermatogonial stem cell maintenance in cultured testes of medaka as a lower vertebrate model. These results demonstrate that RA plays a highly conserved role in germ cell development from fish to mammals.

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Interordinal Chimera Formation Between Medaka and Zebrafish for Analyzing Stem Cell Differentiation

Cited by 31 publications

References 45 publications

Derivation and Long-Term Culture of an Embryonic Stem Cell-Like Line from Zebrafish Blastomeres Under Feeder-Free Condition

Derivation and Long-Term Culture of an Embryonic Stem Cell-Like Line from Zebrafish Blastomeres Under Feeder-Free Condition

Medaka piwi is Essential for Primordial Germ Cell Migration

Retinoic Acid Regulates Germ Gene Transcription In Vitro and Spermatogenesis in Testicular Organ Culture

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