Abstract
In this study, trehalose was investigated for its cryoprotective effects on ovarian granulosa cells (bGCs) of cattle. Five concentrations of trehalose at 0, 0.2, 0.4, 0.6 and 0.8 mol/L were added to the cryopreservation medium of bGCs, and the effects on the quality of frozen-thawed bGCs were assessed. The results indicate that the use of cryopreservation medium containing 0.2 and 0.4 mol/L of trehalose resulted in a greater rate of bGC viability compared to those of other groups (P<0.05). Culturing with trehalose at 0.2 and 0.4 mol/L increased 17β- estradiol (E2)and decreased progesterone (P4)production (P < 0.05) in post-thawed bGCs. Compared with the control group, the intracellular Ca2+ concentrations of frozen-thawed bGCs were less in all treatment groups (P<0.05), and the least Ca2+ concentration was observed in the group containing 0.4 mol/L trehalose. The plasma membrane potentials of frozen-thawed bGCs were greater in the groups with 0.2 and 0.4 mol/L trehalose, and the group treated with 0.4 mol/L trehalose had the greatest membrane potential in comparison to other groups (P < 0.05). The relative abundance of the CYP19 mRNA in frozen-thawed bGCs was greater in the groups containing 0.2, 0.4 and 0.6 mol/L trehalose, and relative abundances of FSHR and BCL2 mRNA were greater in the group of bGCs treated with 0.2 mol/L trehalose (P<0.05). Trehalose treatment at 0.4, 0.6 and 0.8 mol/L had an inhibitory effect on BAX gene transcription in frozen-thawed bGCs (P<0.05). In summary, trehalose exhibited a greater cryoprotective effect on bGCs than basic cryopreservation medium.
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