RT journal article
T1 Establishment and characterisation of single cell-derived embryonic stem cell lines from the gilthead seabream, Sparus aurata
A1 Verges Castillo, Alba
A1 González Vargas, I.A.
A1 Muñoz Cueto, José Antonio
A1 Martín Robles, Águeda Jimena
A1 Pendón Meléndez, Carlos
A2 Biología
A2 BiomedicinaBiotecnología y Salud Pública
K1 Embryonic stem cell lines
K1 Monoclonal cell line
K1 pcna
K1 In vitro studies
K1 Sparus aurata
K1 Embryoid bodies
K1 Aquaculture
AB An important bottleneck in fish aquaculture research is the supply and maintenance of embryos, larvae, juvenileand adult specimens. In this context, cell lines represent alternative experimental models for in vitro studies thatcomplement in vivo assays. This allows us to perform easier experimental design and sampling and avoid thesacrifice of animals. Embryonic stem (ES) cell lines have attracted increasing attention because they have thecapability to proliferate indefinitely and could be differentiated into any cell type of the organism. To minimisecell heterogeneity and increase uniformity of in vitro studies results, in this manuscript we report the developmentand characterisation of two single cell-derived ES cell lines (monoclonal) from the morula stage embryos ofthe gilthead seabream, Sparus aurata, named as SAEC-A3 and SAEC-H7. Both cell lines have been passaged forover 100 times, indicating the establishment of long-term, immortalised ES cell cultures. Sequence analysesconfirmed the seabream origin of the cell lines, and growth analyses evidenced their high viability and proliferatingactivity, particularly in culture medium supplemented with 10–15% fetal bovine serum and 22 ◦C. Bothcell lines showed the ability to generate embryoid bodies and show different sensitivity and response to all-transretinoic acid. The analysis of epithelial (col1α1) and neuronal (sox3) markers in differentiated cultures revealedthat SAEC-A3 tended to differentiate towards epithelial-like cells whereas SAEC-H7 tended to differentiate towardsneuronal-like cells. Both cell lines were efficiently transfected with pDsRed2-ER and/or pEGFP-N1 plasmids,indicating that they could represent useful biotechnological tools. Daily expression of pcna showedsignificant expression rhythms, with maximum levels of cell proliferation during the day-night transition.Currently, these cell lines are being successfully used as experimental models for the study of cellular metabolism,physiology and rhythms as well as for toxicological, pharmacological and gene expression analyses.
PB ELSEVIER
SN 1096-4959
YR 2021
FD 2021-05
LK http://hdl.handle.net/10498/24934
UL http://hdl.handle.net/10498/24934
LA eng
NO This work was funded by grants from the Spanish Ministry of Science,Innovation and Universities (AGL2017-82582-C3-1-R, AGL2013-49027-C3-2-R) to JAMC. AVC is a FPI fellow (PRE2018-085642) fundedby the Spanish Ministry of Science, Innovation and Universities.
DS Repositorio Institucional de la Universidad de Cádiz
RD 07-nov-2026