卵巢皮质组织厚度在人卵巢组织异种移植兔模型中对移植效果的影响

Effects of Graft Thickness on Xenotransplantation of Human Ovarian Tissue in Rabbit Model

目的:研究不同厚度的卵巢组织移植物对人卵巢组织异种移植效果的影响。方法:将妇科手术中取得的人新鲜卵巢皮质组织切割成不同厚度的组织片,并根据厚度分为A组(2 mm×2 mm×2 mm)和B组(2 mm×2 mm×1 mm)。然后将卵巢组织移入去势4周的兔背肌中,构建人卵巢组织异种移植兔模型,每兔移植8片,每组5只动物,移植后6周回收。采用放射性免疫分析法测兔血清雌二醇(E2)含量,HE染色观察移植组织显微镜下形态,CD31染色计数新生血管数,TUNEL标记移植回收卵巢组织凋亡细胞。结果:E2水平B组恢复较快,在移植后的第1、2、4、5周B组较同一时间的A组升高更显著(P<0.05);B组卵巢组织回收率(91.7%)稍高于A组(86.7%),HE染色B组的卵巢基质保存完整,纤维化率低于A组;B组的平均新生血管数(17.7±5.8条)较A组(6.3±2.9条)明显升高(P<0.01),B组中TUNEL染色阳性细胞率明显低于A组(13.45±5.50%vs 45.31±9.29%,P<0.01)。结论:移植物厚度对卵巢移植后的效果有明显影响,厚1 mm的小组织片在人新鲜卵巢组织异种移植兔模型中较为适合。

Objective： To investigate the effect of graft thickness on xenotransplantation of human ovarian tissue. Methods： Fresh human ovarian tissue from patients was divided into two parts （two groups）, group A： one part of tissue was cut into strips of 2 mm × 2 mm × 2 mm, group B： another part of tissue was cut into strips of 2 mm × 2 mm × 1 mm. Then they were transplanted into the back muscle of rabbits until 6 weeks after transplantation. The E2 level in the rabbit serum was analyzed by radioimmunoassay, histological morphology was observed by HE staining under microscope, new vessels were detected by CD31 immunohistochemical staining and apoptotic cells were detected by TUNEL assay. Results： The E2 level in group B rose faster and higher than that in group A at week 1, 2, 4, 5 after transplantation （P〈0.05）, the recovery rate in group B （91.7%） was higher than that in group A （86.7%）, group B had better ovarian stroma and less fibrosis. The average number of new vessels in group B was significantly higher than that in group A （17.7 ±5.8 vs 6.3 ±2.9; P〈0.01）, and the percentage of TUNEL positive-staining cells in group B was significantly lower than that in group A （13.45 ± 5.50% vs 45.31 ±9.29%; P〈0.01）. Conclusion： The thickness of grafts has a great effect on human ovarian tissue xenograft, 1 mm thickness is suitable for the rabbit xenograft model.