Development of vitrified-warmed mouse embryos co-cultured with polarized or non-polarized uterine epithelial cells using sequential culture media.
OBJECTIVE
To our knowledge, little is known about the effect of polarized and non-polarized uterine epithelial cells on cryopreserved embryo growth. This study was, therefore, set up to investigate the effect of these monolayers together with sequential culture media on vitrified-warmed mouse embryos in terms of blastocyst development, blastocyst quality, incidence of apoptosis and related genes expression.


METHODS
Two cell vitrified-warmed mouse embryos were cultured in G-1ver3 medium to the eight-cell stage when they were randomly assigned to three treatment groups of no co-culture (control), non-polarized and polarized mouse uterine epithelial monolayer co-culture. The culture medium was G-2ver3 during the treatment phase. After 96 h on treatment, the significance of differences were evaluated by the one way analysis of variance for continuous data.


RESULTS
In the polarized monolayer group, the hatched blastocyst formation and blastocyst quality improved significantly than other two groups (P < 0.05). Whereas the incidence of apoptosis and related gene expression such as Bax were higher in the blastocysts of control group (P < 0.05). The relative abundance of Bcl-2 mRNA to the beta-tubulin was similar for all treatments.


CONCLUSIONS
Co-culture system involving polarized uterine epithelial cells and sequential culture media is a promising method for the improvement of vitrified-warmed mouse embryo development.
Department of Anatomy, Faculty of Medical Sciences, Tarbiat Modares University, Tehran, Iran.
Mehri
M
Azadbakht
Mojtaba Rezazadeh
MR
Valojerdi
D000818:Animals; D017209:Apoptosis / Q000235:genetics; D002452:Cell Count; D016764:Cell Polarity / Q000187:drug effects* / Q000502:physiology; D002470:Cell Survival; D002478:Cells, Cultured; D018920:Coculture Techniques; D015925:Cryopreservation; D017077:Culture Media, Conditioned / Q000494:pharmacology*; D046149:Embryo Culture Techniques / Q000379:methods*; D004622:Embryo, Mammalian; D047108:Embryonic Development / Q000187:drug effects*; D004847:Epithelial Cells / Q000187:drug effects* / Q000502:physiology; D005260:Female; D006358:Hot Temperature; D051379:Mice; D014599:Uterus / Q000166:cytology / Q000502:physiology*
D017077:Culture Media, Conditioned