Introduction: The comparison of the effects of six different media macromolecule-supplements on development and the cleavage rates of the preimplantation mouse embyros and the selection of the best macromolecule(s).
Materials and Methods: One-cell mouse embryos (NMRI strain) were cultured in T6 and supplemented with one of the following macromolecules: Bovine Serum Albumine (BSA, 4 mg/ml), Hyaluronic Acid (HA, 0.5 mg/ml), Albuminar-5 (10%), Fetal Calf Serum (FCS, 10%), Human Follicular Fluid (HFF, 10%) and Polyvinyl Alcohol (PVA, 0.1mg/ml). All the treatments were provided with ethylenediamine tetraacetic acid (EDTA, 0.02mM). Every 24 hour, all embryos were scored in each treatment to define the stage of the development which was attained during the five days following the treatment. By staining the embryos with ethidium bromide, the number of the cells in each embryos was counted. The blastocyst development data was evaluated by chi-square test while the total number of the cells in each blastocyst were analyzed using Tukey-Kramer multiple comparison test.
Results: After 120h, embryos from the treatments that included BSA, HA, PVA and HFF had a significantly higher blastocyst and blastocyst cell number, when compared to embryos developed in FCS, Albuminar-5 and/or free-macromolecule treatments. [blastocyst: (P<0.0001) and blastocyst cell number: (at least P<0.05)]. While, comparison of developmental and cleavage rates in BSA, HA, PVA and HFF did not exhibit difference [blastocyst: (63-67%) and blastocyst cell number (120?7)-(131?7)].
Conclusion: The following macromolecules: BSA, HA, HFF, and PVA can support the development of one-cell mouse embryos to the blastocyst stage in vitro, while the other macromolecule (FCS and Albuminar-5) have lower effects. Since HA is a physiological macromolecule which is found in female reproductive tract, therefore, it is more reasonable to be used to supplement the defined media.