LIRANS Institute of Research in the Applied Natural Sciences, University of Bedfordshire, Luton, UK.
Theriogenology. 2010 Jul 15;74(2):290-303. doi: 10.1016/j.theriogenology.2010.02.013. Epub 2010 May 8.
There have been no reported methods for in vitro growth of early stage ovarian follicles for fish and their cryopreservation is still under investigation. If cryopreservation of early stage ovarian follicles can be achieved, in vitro procedures for ovarian follicle culture, development, ovulation and fertilisation after cryopreservation would be needed. The aim of the present study was to develop an in vitro culture method for early stage zebrafish ovarian follicles for use after their cryopreservation. Procedures for in vitro culture of stage I (primary growth) and stage II (cortical alveolus) ovarian follicles were developed. The effects of concentration of L-15 medium, pH and the concentration of human chorionic gonadotropin (hCG) and activin A were studied. The results demonstrated that early stage zebrafish ovarian follicles can be cultured in vitro for 24 h, stage I and II ovarian follicles can grow to the sizes of early stage II and stage III ovarian follicles after hCG treatment. The method developed here is effective for assessing early stage zebrafish ovarian follicles growth competence in vitro. The results from the present study indicated that in vitro culture is the most reliable method for assessing ovarian follicle viability when compared with vital dye staining methods.
目前尚无鱼类早期卵巢滤泡体外生长的报道方法，其冷冻保存仍在研究中。如果能够实现早期卵巢滤泡的冷冻保存，就需要进行卵巢滤泡培养、发育、排卵和冷冻保存后的受精的体外程序。本研究的目的是开发一种用于冷冻保存后使用的斑马鱼早期卵巢滤泡的体外培养方法。建立了体外培养 I 期（初级生长）和 II 期（皮质小泡）卵巢滤泡的方法。研究了 L-15 培养基浓度、pH 值以及人绒毛膜促性腺激素（hCG）和激活素 A 的浓度对卵巢滤泡生长的影响。结果表明，早期斑马鱼卵巢滤泡可以在体外培养 24 小时，经 hCG 处理后，I 期和 II 期卵巢滤泡可以生长到早期 II 期和 III 期卵巢滤泡的大小。本研究建立的方法可有效评估早期斑马鱼卵巢滤泡的体外生长能力。与活染料染色方法相比，本研究结果表明，体外培养是评估卵巢滤泡活力最可靠的方法。
