Volume 16, Issue 51 (Jul 2006)
J Mazandaran Univ Med Sci 2006, 16(51): 6-19 |
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Baghban Ëslami Nezhad M, Rezazadeh M, Kazemi S, Ëftekhari Yazdani P. The effects of human oviduct polarized, non-polarized monolayer epithelial cells on the formation of mouce two-cell embryo . J Mazandaran Univ Med Sci 2006; 16 (51) :6-19
URL: http://jmums.mazums.ac.ir/article-1-105-en.html
URL: http://jmums.mazums.ac.ir/article-1-105-en.html
Abstract: (12137 Views)
Background and purpose: Ëmbryo co-culture systems with somatic cells have been a subject of extensive study as valuable tools to promote embryo in vitro development. The aim of this study was to evaluate, the Ëffects of a special kind of co-culture system in which somatic cells were cultured as polarized monolayer on mouse embryo development.
Materials and methods : Human oviduct was obtained from patients who had undergone total abdominal hysterectomy. The epithelial cells were isolated and cultured either on ËÇM (Ëxtra-cellular matrix) gel or plastic surfaces as polarized or non polarized monolayer respectively. Ëpithelial nature of cultured cells was confirmed by immunocytochemistry and the polarized condition was evaluated using transmission electron microscopy. Two cell NMRÏ mouse embryos were flushed and cultured on polarized, non-polarized monolayer and medium alone. The rate of development in all groups were daily determined and statistically compared. Ât the end of the cultivation period, trophoectodam (TË) and inner cell mass (ÏÇM) were stained differentially and the number of blastocyst cells were counted and compared statistically.
Results : Çultured cells were epithelial in nature and those cultured on ËÇM gel had highly columnar polarized appearance, with tight junction between adjacent cells. The cells cultured on plastic were spindle shape in sections and there was no evidence of tight junction between cells. The co-culture results showed that during cultivation period, polarized cells improved embryos in vitro development more than polarized monolayer in terms of both blastocyst formation rate and cellularity with statistically significant differences.
Çonclusion: Polarized oviductal epithelial cells have positive effects on embryo development compared to non-polarized cells. These effects appear as high cellular blastocyst in co-culture systems.
Materials and methods : Human oviduct was obtained from patients who had undergone total abdominal hysterectomy. The epithelial cells were isolated and cultured either on ËÇM (Ëxtra-cellular matrix) gel or plastic surfaces as polarized or non polarized monolayer respectively. Ëpithelial nature of cultured cells was confirmed by immunocytochemistry and the polarized condition was evaluated using transmission electron microscopy. Two cell NMRÏ mouse embryos were flushed and cultured on polarized, non-polarized monolayer and medium alone. The rate of development in all groups were daily determined and statistically compared. Ât the end of the cultivation period, trophoectodam (TË) and inner cell mass (ÏÇM) were stained differentially and the number of blastocyst cells were counted and compared statistically.
Results : Çultured cells were epithelial in nature and those cultured on ËÇM gel had highly columnar polarized appearance, with tight junction between adjacent cells. The cells cultured on plastic were spindle shape in sections and there was no evidence of tight junction between cells. The co-culture results showed that during cultivation period, polarized cells improved embryos in vitro development more than polarized monolayer in terms of both blastocyst formation rate and cellularity with statistically significant differences.
Çonclusion: Polarized oviductal epithelial cells have positive effects on embryo development compared to non-polarized cells. These effects appear as high cellular blastocyst in co-culture systems.
Type of Study: Research(Original) |
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