Effects of fetal bovine serum on trophectoderm and primitive endoderm cell allocation of in vitro-produced bovine embryos

Author(s)

Luedke, Felipe Eduardo ; da Costa, Caroline Pereira ; Mendes, Camilla Mota ; dos Santos Hamilton, Thais Rose ; Milazzotto, Marcella Pecora ; Assumpcao, Mayra E. O. A. ; Goissis, Marcelo Demarchi

• Document type: Journal article
• Source: ZYGOTE; v. 31, n. 1, p. 7-pg., 2022-10-24.

Abstract

Supplementing embryonic culture medium with fetal bovine serum (FBS) renders this medium undefined. Glucose and growth factors present in FBS may affect the results of cell differentiation studies. This study tested the hypothesis that FBS supplementation during in vitro culture (IVC) alters cell differentiation in early bovine embryo development. Bovine embryos were produced in vitro and randomly distributed into three experimental groups at 90 h post insemination (90 hpi): the KSOM-FBS group, which consisted of a 5% (v/v) FBS supplementation; the KSOM33 group, with the renewal of 33% of medium volume; and the KSOM-Zero group, without FBS supplementation nor renewal of the culture medium. The results showed that the blastocyst rate (blastocyst/oocytes) at 210 hpi in the KSOM-FBS group was higher than in the KSOM-Zero group but not different from the KSOM33 group. There were no significant changes in metabolism-related aspects, such as fluorescence intensities of CellROX Green and MitoTracker Red or reduced nicotinamide adenine dinucleotide (NADH) and flavin adenine dinucleotide (FAD+). Immunofluorescence analysis of CDX2 revealed that the lack of FBS or medium supplementation reduced the number of trophectoderm (TE) cells and total cells. Immunofluorescence analysis revealed a reduction of SOX17-positive cell numbers after FBS supplementation compared with the KSOM33 group. Therefore, we concluded that FBS absence reduced blastocyst rates; however, no reduction occurred when there was a 33% volume renewal of the medium at 90 hpi. We also concluded that FBS supplementation altered TE and primitive endoderm cell allocation during early bovine embryo development. (AU)

• FAPESP's process: 18/18924-8 - Influence of the HIPPO signaling pathway on the segregation between the inner cell mass and the trophectoderm in bovine embryos
• Grantee: Caroline Pereira da Costa
• Support Opportunities: Scholarships in Brazil - Master
• FAPESP's process: 17/09576-3 - Studies on cell differentiation mechanisms during early bovine embryo development
• Grantee: Marcelo Demarchi Goissis
• Support Opportunities: Research Grants - Young Investigators Grants
• FAPESP's process: 19/03014-9 - Insertion of fluorescent reporter gene in Sox2 gene region to study cell differentiation in early bovine embryo development
• Grantee: Felipe Eduardo Luedke
• Support Opportunities: Scholarships in Brazil - Master
• FAPESP's process: 17/25574-0 - Studies on cell differentiation mechanisms during early bovine embryo development
• Grantee: Marcelo Demarchi Goissis
• Support Opportunities: Scholarships in Brazil - Young Researchers