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使用新型胚胎玻璃化试剂盒成功玻璃化化驼胚胎。

Successful vitrification of dromedary camel embryos using a novel embryo vitrification kit.

机构信息

Camel Reproduction Centre, PO Box 79914, Dubai, United Arab Emirates.

Camel Reproduction Centre, PO Box 79914, Dubai, United Arab Emirates; Cria Genesis, Ocean Grove, Victoria, Australia.

出版信息

Anim Reprod Sci. 2020 Jul;218:106483. doi: 10.1016/j.anireprosci.2020.106483. Epub 2020 May 6.

Abstract

Embryos (n = 87) collected 8 days after mating and 7 days after ovulation were vitrified using a camel-specific vitrification kit. Vitrification solutions (VS) containing 20% foetal calf serum, with or without 2% bovine serum albumin (BSA) were used to cryopreserve embryos, in three steps VS1 (5 min), VS2 (5 min) and VS3 (30-35 s) at room temperature (RT) before being loaded into open pulled straws and immediately frozen in liquid nitrogen. Embryos were subsequently thawed in warming solutions (WS) in three steps: WS1 at 37 °C (1 min), WS2 at RT (5 min) then into holding media at RT (5-60 min) prior to transfer, in pairs, into recipient camels 6 days after ovulation. There were 42 of 43 embryos viable after vitrification in media without BSA and these were subsequently transferred into 21 recipient females which resulted in ten pregnancies 60 days after transfer (48% pregnancy rate). There were 38 of 44 viable embryos vitrified in media containing BSA that were transferred in pairs into 19 recipient females which resulted in five pregnancies 60 days after transfer (26% pregnancy rate; P > 0.05). Of the total 15 foetuses that developed to 60 days of gestation after vitrification, 11 resulted from embryos of 200-499 μm diameter and four from embryos of 500-700 μm diameter (P > 0.05). There were encouraging results with use of this novel vitrification kit for the commercial application of cryopreservation of camel embryos with a diameter of 300-550 μm.

摘要

将 8 天配种后和 7 天排卵后的胚胎使用骆驼专用的玻璃化液试剂盒进行玻璃化处理。含有 20%胎牛血清的玻璃化溶液(VS),无论是否含有 2%牛血清白蛋白(BSA),用于在室温(RT)下将胚胎在三个步骤 VS1(5 分钟)、VS2(5 分钟)和 VS3(30-35 秒)中进行冷冻保存,然后在装入开放式拉制 straws 并立即在液氮中冷冻。胚胎随后在三个步骤的解冻溶液(WS)中解冻:WS1 在 37°C(1 分钟)、WS2 在 RT(5 分钟),然后在 RT(5-60 分钟)进入保持培养基中,然后成对转移到排卵后 6 天的受体骆驼中。在不含 BSA 的培养基中进行玻璃化处理的 43 个胚胎中有 42 个是存活的,随后将这些胚胎转移到 21 个受体雌性骆驼中,导致 10 个妊娠在转移后 60 天(48%的妊娠率)。在含有 BSA 的培养基中进行玻璃化处理的 44 个胚胎中有 38 个是存活的,然后将这些胚胎成对转移到 19 个受体雌性骆驼中,导致 5 个妊娠在转移后 60 天(26%的妊娠率;P>0.05)。在玻璃化处理后发育到 60 天妊娠的 15 个胎儿中,11 个来自直径为 200-499μm 的胚胎,4 个来自直径为 500-700μm 的胚胎(P>0.05)。使用这种新型玻璃化试剂盒进行商业应用,对直径为 300-550μm 的骆驼胚胎进行冷冻保存,取得了令人鼓舞的结果。

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