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猪胚胎培养。

Culture of pig embryos.

作者信息

Petters R M, Wells K D

机构信息

Department of Animal Science, North Carolina State University, Raleigh 27695-7621.

出版信息

J Reprod Fertil Suppl. 1993;48:61-73.

PMID:8145215
Abstract

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.

摘要

猪胚胎可以采用多种不同策略进行培养,包括一些复杂方法,如在代孕输卵管(兔、羊、小鼠)中进行体内培养、在器官培养中于小鼠输卵管内培养、胚胎与细胞共培养,以及一些简单方法,如在限定培养基或盐溶液中培养。在需要囊胚发育和孵化的情况下,向培养基中添加血清尤为重要。猪的孕体(第10 - 15天)、胚胎盘或源自孕体的细胞系可以在复杂培养基中培养,如含有血清的伊格尔最低限度基本培养基或杜尔贝科改良伊格尔培养基,不过胚胎盘可以在无血清条件下培养。相比之下,早期猪胚胎(单细胞到囊胚)最好在更简单的培养基中培养,例如用于小鼠胚胎的培养基。所使用的培养基在成分上都相对相似。它们含有盐类以及一种或多种能量来源,如葡萄糖、乳酸或丙酮酸,并以牛血清白蛋白作为大分子成分。早期培养猪胚胎的尝试不太成功,但一些胚胎确实发育到了囊胚阶段。最近的报告表明,在培养基成分相对变化不大或没有变化的情况下,发育率要高得多。一些研究人员报告称,在缺乏葡萄糖的培养基中发育有所改善,这与仓鼠等实验动物的研究结果一致。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来源。向培养基中添加牛磺酸和次牛磺酸可提高猪胚胎的体外发育率。我们建议,在可能的情况下,采用一种不同实验室甚至不同物种都能使用的标准培养基。使用一种培养基培养不同物种将简化实验方案,加强对胚胎比较生理学和代谢的研究,并加快不同物种间信息的传递。

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