The establishment of hypothermic mechanical perfusion model of rat ovary in vitro

doi:10.3877/cma.j.issn.1674-3903.2020.04.007

Abstract

Abstract:

Objective

To establish a hypothermic mechanical perfusion (HMP) model of rat ovary in vitro and test its effectiveness.

Methods

26 female SD rats aged 8-10 weeks were divided into donor group, recipient group, normal group and ovariectomy group, with 6 rats in each group. Ovarian tissue of 2 rats was harvested for test after HMP. Ovaries and kidneys from donor group and HMP management only group were placed in Ringer′s solution at 0-4 ℃ for 2 h, and follwed by HMP for 2 h. Ovarian tissue from donor group was transplanted under the recipient′s left kidney capsule after HMP, and the serum anti-Mullerian hormone (AMH) in recipient rat was tested 7 days after transplantation, and the ovaries were harvested for histological analysis, and immunohistochemistry for CD31 and Ki67. ANOVA was used to compare the serum AMH levels in the normal group, the ovariectomy group and the recipient group. The t-test was used to compare the follicle counts and the percentages of Ki67 and CD31 positive cells in the normal group and the recipient group. P<0.05 was considered statistically significant.

Results

No obvious dilatation was observed in the blood vessels of ovarian tissue after HMP. The follicle structure of rats from recipient group was complete 7 days after transplantation, and the number of mature follicles was (7.0±0.6), which was lower than that of the normal group (26.3±3.4), the difference had statistical significance (t=5.633, P<0.05). The serum AMH of recipient group was (4.63±0.87) ng/mL, higher than that of the ovariectomy group [(2.55±0.16) ng/mL], and lower than that of the normal group [(7.39±1.04) ng/mL]. The percentage of Ki67 and CD31 positive cells in ovarian tissue from recipient group 7 days after transplantation were (29.0±2.4)% and (12.3±1.0)%, which had no significant difference with normal group [(21.9±3.8%) and (12.6±1.2%)] (t=1.634 and 0.178, P all>0.05).

Conclusions

The method to construct the model of HMP in vitro of rat ovary in this study is simple and can maintain the normal function of ovary for a short time, providing a new method for organ preservation of ovary.

Key words: Rats, Ovarian transplantation, Hypothermic mechanical perfusion, Ovary

Shichen Zhang, Hanlin Yao, Zhijian Yang, Qifa Ye, Yanfeng Wang. The establishment of hypothermic mechanical perfusion model of rat ovary in vitro[J]. Chinese Journal of Transplantation(Electronic Edition), 2020, 14(04): 229-233.

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References 22

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