CN103881909A - 哺乳动物胚胎毛细管体外培养装置 - Google Patents

哺乳动物胚胎毛细管体外培养装置 Download PDF

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CN103881909A
CN103881909A CN201310267895.0A CN201310267895A CN103881909A CN 103881909 A CN103881909 A CN 103881909A CN 201310267895 A CN201310267895 A CN 201310267895A CN 103881909 A CN103881909 A CN 103881909A
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李相运
孙芳园
张振南
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Hebei Agricultural University
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Abstract

一种哺乳动物胚胎毛细管体外培养装置。它是利用一种能够产生负压的装置将胚胎吸入塑料毛细管内，再将塑料毛细管置入旋转的液体中在培养箱中进行水浴培养，旋转的液体使塑料毛细管产生摆动和震动，不但能够模拟生殖道对胚胎发育的刺激作用，而且显著减小了培养基体积，胚胎质量和发育能力显著提高，而且该装置操作简便，节约耗材，不需要昂贵的培养皿和矿物油。

Description

哺乳动物胚胎毛细管体外培养装置
技术领域
本发明涉及动物胚胎工程领域，特别是涉及一种哺乳动物胚胎体外培养装置，利用该装置可显著提高胚胎体外发育能力。
背景技术
胚胎体外培养是胚胎工程领域的主要技术，在所有的有关胚胎研究领域，如体外受精、试管婴儿、胚胎移植、动物克隆、转基因动物、胚胎干细胞、嵌合体动物等都涉及胚胎体外培养环节，胚胎质量的优劣以及发育能力是影响上述这些研究的主要因素。对胚胎质量和发育能力起决定性作用的是胚胎体外培养条件，包括培养基的组成和培养体系。在过去的30年，培养基的组成已经发生了很大的变化，而培养体系一直沿用传统的微滴培养法，即在培养皿底部做一小液滴，将多个胚胎置于其中，然后在液滴上方覆盖矿物油，最后将培养皿置于二氧化碳培养箱中即可。微滴培养法至少存在两种缺陷，一是无法模拟胚胎在体内发育过程中所受到的各种生殖道刺激，如输卵管蠕动对胚胎的刺激作用。二是微滴的体积显著大于胚胎在体内所处的空间体积，大量培养基不利于胚胎体外发育。这两种缺陷是导致目前胚胎体外培养效率差的重要原因，胚胎质量显著低于体内正常发育的胚胎，因此，我们发明了一种胚胎毛细管体外培养装置，该装置不但能够模拟生殖道对胚胎发育的刺激作用，而且显著减小了培养基体积，胚胎质量和发育能力显著提高，同时，该装置操作简便，节约耗材，不需要昂贵的培养皿和矿物油。通过对国内外专利文献、期刊杂志及其他公开发表的文献（如互联网）进行检索，本发明在国内外未见报道，我们也未公开发表涉及本发明内容的文章，本发明在国内外公众未知。
发明内容
为了克服现有的胚胎体外培养技术的缺陷，提高胚胎培养质量和胚胎发育能力，本发明提供了一种哺乳动物胚胎毛细管体外培养装置。本发明所采用的技术方案是：将塑料毛细管与塑料接头相连接，连接部位用热缩管密封，塑料接头与移液器相匹配，塑料毛细管透明光滑、体积小、耐高温、耐低温、耐腐蚀、高润滑、不粘附、无毒害。塑料毛细管摩擦系数小，表面张力小，无毛细现象，内径狭小，所装培养基体积小。将移液器与塑料接头相连，在解剖显微镜下通过移液器产生的负压将胚胎和培养基吸入塑料毛细管，然后将塑料毛细管置入旋转的液体中，液体旋转的动力来自一种磁力搅拌装置，旋转的液体使塑料毛细管产生摆动和震动，模拟子宫和输卵管的运动环境。最后将整个装置放入二氧化碳培养箱即可。塑料毛细管处于大量旋转的液体中，液体温度恒定，不易受外界环境影响。哺乳动物胚胎毛细管体外培养装置除用于胚胎体外培养外，还可用于体外受精、嵌合体制备、类胚体培养、细胞聚合等研究。与现有技术相比，本发明的有益效果是：提高了哺乳动物胚胎体外培养效率，提高了胚胎质量和发育能力，显著优于现有的微滴培养法，而且该哺乳动物胚胎毛细管体外培养装置节约耗材，不需要昂贵的细胞培养皿和矿物油，操作简便，不易污染。
具体实施方式
将4厘米长、内径0.3毫米的聚全氟乙烯丙烯共聚物塑料毛细管与10微升的聚丙烯塑料接头（与2微升的移液器相匹配）相连接，并用热缩管将连接之处密封，将连接好的塑料毛细管高压灭菌后烘干备用。收集小鼠受精卵或2-细胞胚胎，置于培养基中，将移液器与塑料接头连接，在解剖显微镜下，利用移液器产生的负压将10个受精卵或2-细胞胚胎吸入塑料毛细管，然后将装有胚胎的塑料毛细管置于盛有400毫升蒸馏水的烧杯中，将烧杯放在磁力搅拌器上，之后将整个装置放入二氧化碳培养箱中培养2-4天，培养完成后再用移液器将胚胎取出，用蒸馏水反复冲洗塑料毛细管并高压灭菌后烘干，下次继续使用。

Claims (5)

1.一种哺乳动物胚胎毛细管体外培养装置，塑料毛细管与塑料接头相连接，连接部位用热缩管密封，塑料接头与移液器相匹配，塑料毛细管置于旋转的液体中，其特征是，塑料毛细管透明光滑、体积小、耐高温、耐低温、耐腐蚀、高润滑、不粘附、无毒害，塑料毛细管处于旋转的液体中，液体温度恒定。
2.根据权利要求1所述的哺乳动物胚胎毛细管体外培养装置，其特征是：塑料毛细管摩擦系数小，表面张力小，无毛细现象，内径狭小，所装培养基体积小，利用移液器产生的负压将胚胎吸入塑料毛细管。
3.根据权利要求1所述的哺乳动物胚胎毛细管体外培养装置，其特征是：塑料毛细管处于旋转的液体中，液体旋转的动力来自一种磁力搅拌装置，旋转的液体使塑料毛细管产生摆动和震动。
4.根据权利要求1所述的哺乳动物胚胎毛细管体外培养装置，其特征是：旋转的液体置于二氧化碳培养箱中，液体温度恒定。
5.根据权利要求1所述的哺乳动物胚胎毛细管体外培养装置，其特征是：哺乳动物胚胎毛细管体外培养装置除用于胚胎体外培养外，还可用于体外受精、嵌合体制备、类胚体培养、细胞聚合等研究。
CN201310267895.0A 2013-07-01 2013-07-01 哺乳动物胚胎毛细管体外培养装置 Pending CN103881909A (zh)

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Publication number Priority date Publication date Assignee Title
CN106701528A (zh) * 2015-08-20 2017-05-24 陈薪 快捷挪卵/胚胎装置

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Cited By (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN106701528A (zh) * 2015-08-20 2017-05-24 陈薪 快捷挪卵/胚胎装置

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Application publication date: 20140625