Isolation and establishment of a new embryonic-like stem cell line from zebrafish

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Date
2014
Authors
Youn Sing, Lee
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Embryonic stem (ES) cells established from various fish species using feederfree method had been reported in many cases. However, zebrafish ES-like cell lines were currently limited and required the addition of fish embryo extract (FEE) in medium formulation. FEE was not easily available to laboratories that lack of aquatic housing system. Furthermore, it may raise ethical issues as it involved destruction of large numbers of developing fish embryos to support the growth of in vitro cells. Therefore, the aim of this study was to derive an ES-like cell line from zebrafish blastomeres without the use of feeder cells and FEE. In this study, a new cell line, ZES4 was successfully isolated and maintained in ES-like cell state and showed in vitro differentiation potentials upon changing the culture conditions. Differentiated cells were identified through morphological and immunocytochemistry using cell lineage specific markers. However, these cell lineage specific markers were also found in undifferentiated ZES4 cells. Stage specific embryonic antigen 3 (SSEA3) and SSEA4 was detected in both blastula stage embryos and undifferentiated ZES4 cells. SSEA1 was detected in minority of ZES4 cells in high density differentiation culture conditions after 40 days. GFP-labeled ZES4 cells were able to contribute to chimera formation in somite, near gonad region and functional heart of the hosts. In conclusion, ZES4 has the ability to differentiate in vitro and in vivo which is comparable to other fish ES cells.
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