Development and Characterization of Embryonic Stemlike Cell Culture from Carassius auratas (Linnaeus, 1758)

Abstract

Embryonic stem cells are pluripotent cells, capable
of differentiating into various cell types. In the
present study, feeder cell-free culture of midblastula
embryos of the Goldfish (Carassius auratus)
was carried out. Embryonic stem cells were derived
from mid-blastula stage embryos using pronase to
release the inner cell mass (ICM). The cells were
cultured in Leibovitz-15 medium on gelatin-coated
cell culture well plates supplemented with 15% fetal
bovine serum, 1% goldfish serum, basic fibroblast
growth factor and leukaemia inhibitory factors (LIF)
and were incubated at 28°C. The ES-like cells were
characterized by their unique round and polygonal
morphology, elevated activity of alkaline phosphatase,
spherical three-dimensional embryoid body
formation, chromosomal diploid number (2n=50).
The expression of Oct4, a member Oct family of
POU transcription factors, which play a key role in
regulating stem cell pluripotency and differentiation,
was used for characterization of undifferentiated
pluripotent ES cells. The expression of Oct4 in
embryonic stem-like cell (2nd passage) and gonads
of goldfish was studied by RT- PCR. Embryonic
stem-like cells culture from the mid blastula embryo
of C. auratus would be a useful tool for cellular
development and differentiation studies.