猪胚胎的培养。

R. Petters, K. Wells
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引用次数: 926

摘要

猪胚胎的培养可以采用多种不同的策略,包括复杂的方法,如在代孕输卵管(兔、羊、小鼠)中进行体内培养,在小鼠输卵管中进行器官培养,以及胚胎与细胞的共培养,此外还有简单的方法,如在特定培养基或盐溶液中培养。在需要胚泡发育和孵化的培养基中加入血清是特别重要的。猪胚(10-15天)、胚盘或由胚盘衍生的细胞系可以在复杂的培养基中培养,如Eagle的最低基本培养基或Dulbecco的改良Eagle培养基中添加血清,尽管胚盘可以在没有血清的情况下培养。相比之下,早期猪胚胎(单细胞到囊胚)最好在简单的培养基中培养,如用于小鼠胚胎的培养基。所使用的介质在组成上都是相对相似的。它们含有盐和一种或多种能量来源,如葡萄糖、乳酸或丙酮酸,并以牛血清白蛋白为大分子成分。早期培养猪胚胎的尝试并不十分成功,但一些胚胎确实发育到了囊胚阶段。最近的报告显示,发展速度要高得多,而媒介构成的变化相对很少或没有变化。一些工作人员报告说,在缺乏葡萄糖的培养基中,发育得到了改善,这与在仓鼠等实验动物身上的发现是一致的。在缺乏葡萄糖、乳酸和丙酮酸的培养基中,谷氨酰胺可以作为唯一的外源能量。在培养基中添加牛磺酸和次牛磺酸可以促进猪胚的体外发育。我们建议,在可能的情况下,采用一种标准培养基,可以用于不同的实验室,也许,不同的物种。对不同物种使用同一种培养基将简化实验方案,加强比较胚胎生理学和代谢的研究,并加快在不同物种中获得的信息的传递。
本文章由计算机程序翻译,如有差异,请以英文原文为准。
Culture of pig embryos.
Pig embryos can be cultured using a number of different strategies including complex approaches like culture in vivo in a surrogate oviduct (rabbit, sheep, mouse), culture in mouse oviducts in organ culture, and co-culture of embryos with cells in addition to simple approaches like culture in defined media or salt solutions. Addition of serum to medium has been of particular importance where blastocyst development and hatching are required. Pig conceptuses (day 10-15), embryonic discs or cell lines derived from conceptuses can be cultured in complex media like Eagle's minimal essential medium or Dulbecco's modified Eagle's medium with serum, although embryonic discs can be cultured in the absence of serum. In contrast, early stage pig embryos (one-cell to blastocyst) are best cultured in simpler media such as those used for mouse embryos. The media that have been used are all relatively similar in composition. They contain salts and one or more energy sources such as glucose, lactate, or pyruvate with BSA as a macromolecular component. Early attempts to culture pig embryos were not very successful, but some embryos did develop to the blastocyst stage. More recent reports indicate a much higher rate of development with relatively little or no change in media composition. Some workers have reported improved development in medium lacking glucose, which is consistent with findings with laboratory animals such as hamsters. Glutamine can serve as the sole exogenous energy source in medium lacking glucose, lactate and pyruvate. Addition of taurine and hypotaurine to culture medium enhances development of pig embryos in vitro. We suggest, where possible, adoption of a standard medium that could be used by different laboratories and, perhaps, with different species. Use of one medium for different species would simplify experimental protocols, enhance studies of comparative embryonic physiology and metabolism, and expedite transfer of information obtained in different species.
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