Although cryopreservation of ovarian tissue has advanced greatly, it remains a challenge, and protocols should be optimized to handle the heterogeneous nature of ovarian samples. In an effort to address this factor, the present study evaluated the effects of corpus luteum (CL) and side of ovaries (right versus left) on cellular morphology and viability of vitrified bovine ovarian fragments in a closed system. The ovaries were categorized according to whether they had a CL and which side they were on, and then divided into six groups: 1) CL+ (with CL) group; 2) CL− (without CL) group; 3) right ovaries group; 4) left ovaries group;5) fresh control group (ovaries without vitrification or culture that were not selected for CL or ovarian side) and 6) In vitro culture medium control group (non-vitrified ovaries that were not selected for the presence or absence of CL or side of the ovaries). The ovarian tissue Cryosystem was used to vitrify the ovarian fragments. Ovarian tissues were intended for viability and histological tests, and immunohistochemistry was performed to immunolocalize Aquaporin 3 and connexin 43 and 37. The current study shows that the CL− and right groups had the greatest percentage of follicles with normal morphology compared to other vitrified-warmed groups. Furthermore, the levels of necrosis and tissue damage of the right cultured group were the lowest compared to other groups. For the first time, the present study suggests that bovine ovarian tissue vitrification can be improved by considering the origin of the ovaries.