Enhanced in vitro maturation of canine oocytes by oviduct epithelial cell co-culture

Oviduct flushing is enriched by a wide variety of nutrients that guide the 3–4 days journey of pre-implantation embryo through the oviduct as it develops into a competent blastocyst (BL). However, little is known about the specific requirement and role of these nutrients that orchestrate the early stages of embryonic development. In this study, we aimed to characterize the effect of in vitro-derived bovine oviduct epithelial cell (BOECs) secretion that mimics the in vivo oviduct micro-fluid like environment, which allows successful embryonic development. In this study, the addition of an in vitro derived BOECs-condition media (CM) and its isolated exosomes (Exo) significantly enhances the quality and development of BL, while the hatching ability of BLs was found to be high (48.8%) in the BOECs-Exo supplemented group. Surprisingly, BOECs-Exo have a dynamic effect on modulating the embryonic metabolism by restoring the pyruvate flux into TCA-cycle. Our analysis reveals that Exo treatment significantly upregulates the pyruvate dehydrogenase (PDH) and glutamate dehydrogenase (GLUD1) expression, required for metabolic fine-tuning of the TCA-cycle in the developing embryos. Exo treatment increases the influx into TCA-cycle by strongly suppressing the PDH and GLUD1 upstream inhibitors, i.e., PDK4 and SIRT4. Improvement of TCA-cycle function was further accompanied by higher metabolic activity of mitochondria in BOECs-CM and Exo in vitro embryos. Our study uncovered, for the first time, the possible mechanism of BOECs-derived secretion in re-establishing the TCA-cycle flux by the utilization of available nutrients and highlighted the importance of pyruvate in supporting bovine in vitro embryonic development.

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Improved bovine embryo production in an oviduct-on-a-chip system: prevention of poly-spermic fertilization and parthenogenic activation

Lab on a Chip ◽

10.1039/c6lc01566b ◽

2017 ◽

Vol 17 (5) ◽

pp. 905-916 ◽

Cited By ~ 21

Author(s):

Marcia A. M. M. Ferraz ◽

Heiko H. W. Henning ◽

Pedro F. Costa ◽

Jos Malda ◽

Ferry P. Melchels ◽

...

Keyword(s):

Epithelial Cell ◽

3D Printing ◽

Cell Cultures ◽

Bovine Embryo ◽

Embryo Production ◽

Epithelial Cell Cultures ◽

Oviduct Epithelial Cell

The use of 3D-printing in bovine oviduct epithelial cell cultures allows better bio-mimicking of embryo production than classical in vitro fertilization.

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The interaction of fresh and frozen-thawed ram spermatozoa with oviducal epithelial cells in vitro

Reproduction Fertility and Development ◽

10.1071/rd00032 ◽

2000 ◽

Vol 12 (6) ◽

pp. 237 ◽

Cited By ~ 11

Author(s):

L. Gillan ◽

G. Evans ◽

W. M. C. Maxwell

Keyword(s):

Epithelial Cell ◽

Epithelial Cells ◽

Small Proportion ◽

Oestrous Cycle ◽

Cell Monolayers ◽

Oviduct Epithelial Cell ◽

Epithelial Cell Monolayers ◽

Oviduct Epithelial Cells

In order to investigate the interaction of fresh and frozen–thawed spermatozoa with oviduct epithelial cells, spermatozoa were co-incubated with ovine oviduct epithelial cell monolayers (OECM) derived from either complete oviducts, at any stage of the oestrous cycle (Experiments 1 and 2), or from different regions of the oviduct at different stages of the cycle (Experiment 3). Fresh and frozen—thawed spermatozoa displayed different patterns of binding to, and release from, the OECM. Frozen—thawed spermatozoa immediately bound to the complete oviduct OECM and were released after 2 h. A small proportion of fresh spermatozoa bound immediately, increasing to a maximum after 2 h, and were gradually released thereafter. When only the cells that were released from the OECM were observed by chlortetracycline staining in Experiment 2, it was found that the presence of an OECM increased the number of capacitated fresh spermatozoa while decreasing the number of capacitated frozen–thawed spermatozoa. Overall, the OECM advanced the membrane state of both types of spermatozoa from uncapacitated to acrosome-reacted. Fresh and frozen—thawed spermatozoa bound to OECM derived from the cells of the isthmus and the ampulla in similar proportions. However, more spermatozoa were capacitated when incubated with OECM derived from isthmic rather than ampullary cells. Higher proportions of fresh spermatozoa bound to, and were acrosome-reacted following incubation with OECM derived from post- rather than pre-ovulatory tracts. Such differences were not observed for frozen—thawed spermatozoa. The findings reported in this study show that fresh and frozen—thawed spermatozoa behave differently when in contact with oviduct cells in vitro. This may be a consequence of the more advanced membrane state of the frozen spermatozoa upon thawing.

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