常用实验动物带血管蒂全卵巢灌注及冷冻保存方法

Perfusion and cryopreservation of whole ovaries with vascular pedicles in common experimental animals

目的采用独创的已获专利授权的灌注针和灌注技术,对常用实验动物(成年兔、豚鼠和大鼠)的带血管蒂全卵巢进行灌注,研究可调节灌注针是否适用于不同体型动物全卵巢的灌注,同时采用经典的冷冻方案进行冷冻及解冻。方法分别取6只性成熟雌性中国大耳白兔、6只Dunkan-Hartley系豚鼠、6只SD大鼠各12个带血管蒂全卵巢进行实验。结果计数兔、豚鼠、大鼠3种动物卵巢组织HE染色切片中形态正常的原始卵泡率,结果显示差异均无统计学意义(P>0.05);血管损伤主要在远卵巢端,3种动物近卵巢端的各组动脉、静脉均未见明显损伤。结论通过调节麦管针的粗细可用于灌注大小不同的动物卵巢,初步证实了相同的灌注压力(60 mmHg)和灌注速率(1 ml/min)同时适用于不同体积的兔、豚鼠和大鼠卵巢。

Objective Using the perfusion needle which is authorized and special perfusion technology to perfuse the whole ovary with vascular pedicle of common animals,such as rabbits,guinea pigs and rats,then comparative analysis of the adjustable perfusion needles whether can be applied to different animals,whether this kind of perfusion pressure and perfusion rate is appropriate for different sizes of ovaries,and using the classic cryopreservation protocol to freeze and thaw.Methods Collecting 12 ovaries respectively from 6 chinese sexual maturity rabbits,6 Dunkan-Hartley guinea pigs and 6 SD rats to do experiments.Results Through HE staining to count the normal proportion of primordial follicles in each section,the result of HE staining in three animals indicated that there was no statistical significance(P>0.05),vascular injury was mainly in the upper(far away from ovary),there was no obvious damage in lower(close to ovary)in three animals’each group.Conclusion The experiment confirms that through adjusting straw needle size we can perfuse different size of animal organs and under the condition of the same perfusion pressure(60 mmHg)and rate(1 ml/min)it is suitable for rabbits’ovaries,guinea pigs’ovaries and rats’ovaries at the same time.