RESUMO
Objective: In conventional assisted reproductive technology [ART], oocytes are cultured in static microdrops within Petri dishes that contain vast amounts of media. However, the in vivo environment is dynamic. This study assesses in vitro oocyte maturation through the use of a new microfluidic device. We evaluate oocyte fertilization to the blastocyct stage and their glutathione [GSH] contents in each experimental group
Materials and Methods: In this experimental study, we established a dynamic culture condition. Immature oocytes were harvested from ovaries of Naval Medical Research Institute [NMRI] mice. Oocytes were randomly placed in static [passive] and dynamic [active] in vitro maturation [IVM] culture medium for 24 hours. In vitro matured oocytes underwent fertilization, after which we placed the pronucleus [PN] stage embryos in microdrops and followed their developmental stages to blastocyst formation after 3 days. GSH content of the in vitro matured oocytes was assessed by monochlorobimane [MCB] staining
Results: We observed significantly higher percentages of mature metaphase II oocytes [MII] in the passive and active dynamic culture systems [DCS] compared to the static group [P<0.01]. There were significantly less mean numbers of germinal vesicle [GV] and degenerated oocytes in the passive and active dynamic groups compared to the static group [P<0.01]. Fertilization and blastocyst formation rate in the dynamic systems were statistically significant compared to the static cultures [P<0.01]. There was significantly higher GSH content in dynamically matured oocytes compared to statically matured oocytes [P<0.01]
Conclusion: Dynamic culture for in vitro oocyte maturation improves their developmental competency in comparison with static culture conditions
RESUMO
Amniotic fluid represents rich sources of stem cells that can be used in treatments for a wide range of diseases. Amniotic fluid- stem cells have properties intermediate between embryonic and adult mesenchymal stem cells which make them particularly attractive for cellular regeneration and tissue engineering. Furthermore, scientists are interested in these cells because they come from the amniotic fluid that is routinely discarded after birth. In this review we give a brief introduction of amniotic fluid followed by a description of the cells present within this fluid and aim to summarize the all existing isolation methods, culturing, characterization and application of these cells. Finally, we elaborate on the differentiation and potential for these cells to promote regeneration of various tissue defects, including fetal tissue, the nervous system, heart, lungs, kidneys, bones, and cartilage in the form of table.