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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Abstract
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.