Influence of culture medium and age of bovine blastocysts in established colonies of embryonic stem cells.
The objective of this study was to evaluate the potential of bovine IVF blastocysts at different stages of embryonic development in establishing ESC-like. Furthermore, blastocysts cultured in medium containing (10%) and (2.5%) fetal calf serum (FCS) were compared to determine if the serum concentration during in vitro culture alters the blastocyst's potential to establish ESC-like culture. It was observed that only ICM's from expanded blastocysts adhered to the monolayer (n=160) independent of the concentration of serum used during IVF culture. There were no observable differences in potential to establish ESC-like in embryos cultured with 2,5% or 10% FCS . The bFGF didn´t seems to be required for maintenance of bovine ESC-like regardless of culture conditions. Blastocysts and colonies in primary culture and after the first passage were positive for Oct4, Nanog, SSEA-3 and TRA-1-81. Expanded blastocysts gave rise to ESC-like colonies, and the addition of LIF was sufficient to maintain cells undifferentiated in culture.
Departament of Animal Reproduction and Veterinary Radiology (FMVZ)- São Paulo State University-Rubião Jr, s/n 18618-970 Botucatu-SP-Brazil
Midyan Daroz
MD
Guastali
Departament of Animal Reproduction and Veterinary Radiology (FMVZ)- São Paulo State University-Rubião Jr, s/n 18618-970 Botucatu-SP-Brazil
Bianca Andriolo
BA
Monteiro
Departament of Animal Reproduction and Veterinary Radiology (FMVZ)- São Paulo State University-Rubião Jr, s/n 18618-970 Botucatu-SP-Brazil
Rosiara Rosária Dias
RR
Maziero
Departament of Animal Reproduction and Veterinary Radiology (FMVZ)- São Paulo State University-Rubião Jr, s/n 18618-970 Botucatu-SP-Brazil
Daniela Martins
DM
Paschoal
Departament of Animal Reproduction and Veterinary Radiology (FMVZ)- São Paulo State University-Rubião Jr, s/n 18618-970 Botucatu-SP-Brazil
Mateus José
MJ
Sudano
Departament of Animal Reproduction and Veterinary Radiology (FMVZ)- São Paulo State University-Rubião Jr, s/n 18618-970 Botucatu-SP-Brazil
Tatiana da Silva
Tda S
Rascado
Departament of Animal Reproduction and Veterinary Radiology (FMVZ)- São Paulo State University-Rubião Jr, s/n 18618-970 Botucatu-SP-Brazil
João Ferreira Lima
JF
Neto
Luis Eduardo
LE
Vergara
Departament of Animal Reproduction and Veterinary Radiology (FMVZ)- São Paulo State University-Rubião Jr, s/n 18618-970 Botucatu-SP-Brazil
Letícia Ferrrari
LF
Crocomo
In Vitro Brasil Ltda. Rod. SP 340 - KM 166, Mogi Mirim – SP-Brazil
Marc
M
Maserati
Departament of Animal Reproduction and Veterinary Radiology (FMVZ)- São Paulo State University-Rubião Jr, s/n 18618-970 Botucatu-SP-Brazil
Fernanda da Cruz
Fda C
Landim-Alvarenga
D000818:Animals; D001755:Blastocyst / Q000166:cytology* / Q000378:metabolism; D002417:Cattle; D018929:Cell Culture Techniques / Q000379:methods*; D003470:Culture Media / Q000737:chemistry*; D053595:Embryonic Stem Cells / Q000166:cytology*; D005307:Fertilization in Vitro; D005346:Fibroblast Growth Factors / Q000378:metabolism
D003470:Culture Media; D005346:Fibroblast Growth Factors