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Media optimization to promote rat embryonic development to the blastocyst stage in vitro.
Theriogenology ( IF 2.4 ) Pub Date : 2020-07-01 , DOI: 10.1016/j.theriogenology.2020.03.007
Hongsheng Men 1 , Barbara J Stone 2 , Elizabeth C Bryda 1
Affiliation  

Efficient model production in rats that incorporates newly developed genetic editing and embryo transfer tools, such as CRISPR/Cas9 technology and non-surgical embryo transfer, requires availability of an optimal embryo culture system. However, current technologies for in vitro manipulation of rat gametes, including embryo culture techniques, are less advanced compared to those in mice. In this study, we (1) identified a culture medium that was able to support optimal rat embryonic development by comparing two rat culture media: mR1ECM (modified rat 1-cell embryo culture medium) and KSOM-R (modified potassium simplex optimized medium for rats), and (2) evaluated the effect of glutamine dipeptides: alanyl-l-glutamine and glycyl-l-glutamine, on rat embryonic development. We also investigated the possibility of simplifying the KSOM-R culture procedure by increasing the volume of culture medium, reducing the need for daily medium changes. The results showed that rat embryos cultured in KSOM-R developed faster than those cultured in mR1ECM. Both alanyl-l-glutamine and glycyl-l-glutamine showed detrimental effects on rat embryonic development when supplemented in KSOM-R at the same concentration as glutamine. By increasing the volume of KSOM-R, rat zygotes were able to develop without daily medium refreshment at a similar rate and developmental competence as those in smaller volumes with daily medium changes. These results represent important improvements to rat embryo culture methods and will assist in more efficient production of rat models.

中文翻译:

培养基优化促进大鼠胚胎体外发育至囊胚阶段。

结合新开发的基因编辑和胚胎移植工具(如 CRISPR/Cas9 技术和非手术胚胎移植)在大鼠中进行高效模型生产,需要最佳胚胎培养系统的可用性。然而,与小鼠相比,目前用于体外操作大鼠配子的技术,包括胚胎培养技术,还没有那么先进。在本研究中,我们 (1) 通过比较两种大鼠培养基确定了一种能够支持最佳大鼠胚胎发育的培养基:mR1ECM(改良的大鼠 1 细胞胚胎培养基)和 KSOM-R(改良的单一钾优化培养基)大鼠),和 (2) 评估了谷氨酰胺二肽:丙氨酰-l-谷氨酰胺和甘氨酰-l-谷氨酰胺对大鼠胚胎发育的影响。我们还研究了通过增加培养基体积来简化 KSOM-R 培养程序的可能性,减少每天更换培养基的需要。结果表明,在KSOM-R中培养的大鼠胚胎比在mR1ECM中培养的胚胎发育得更快。当以与谷氨酰胺相同的浓度补充 KSOM-R 时,丙氨酰-l-谷氨酰胺和甘氨酰-l-谷氨酰胺都对大鼠胚胎发育产生不利影响。通过增加 KSOM-R 的体积,大鼠受精卵能够在没有每日培养基更新的情况下以与每天更换培养基的较小体积的受精卵相似的速率和发育能力发育。这些结果代表了对大鼠胚胎培养方法的重要改进,并将有助于更有效地生产大鼠模型。减少每天更换培养基的需要。结果表明,在KSOM-R中培养的大鼠胚胎比在mR1ECM中培养的胚胎发育得更快。当以与谷氨酰胺相同的浓度补充 KSOM-R 时,丙氨酰-l-谷氨酰胺和甘氨酰-l-谷氨酰胺都对大鼠胚胎发育产生不利影响。通过增加 KSOM-R 的体积,大鼠受精卵能够在没有每日培养基更新的情况下以与每天更换培养基的较小体积的受精卵相似的速率和发育能力发育。这些结果代表了对大鼠胚胎培养方法的重要改进,并将有助于更有效地生产大鼠模型。减少每天更换培养基的需要。结果表明,在 KSOM-R 中培养的大鼠胚胎比在 mR1ECM 中培养的胚胎发育得更快。当以与谷氨酰胺相同的浓度补充 KSOM-R 时,丙氨酰-l-谷氨酰胺和甘氨酰-l-谷氨酰胺都对大鼠胚胎发育产生不利影响。通过增加 KSOM-R 的体积,大鼠受精卵能够在没有每日培养基更新的情况下以与每天更换培养基的较小体积的受精卵相似的速率和发育能力发育。这些结果代表了对大鼠胚胎培养方法的重要改进,并将有助于更有效地生产大鼠模型。当以与谷氨酰胺相同的浓度补充 KSOM-R 时,丙氨酰-l-谷氨酰胺和甘氨酰-l-谷氨酰胺都对大鼠胚胎发育产生不利影响。通过增加 KSOM-R 的体积,大鼠受精卵能够在没有每日培养基更新的情况下以与每天更换培养基的较小体积的受精卵相似的速率和发育能力发育。这些结果代表了对大鼠胚胎培养方法的重要改进,并将有助于更有效地生产大鼠模型。当以与谷氨酰胺相同的浓度补充 KSOM-R 时,丙氨酰-l-谷氨酰胺和甘氨酰-l-谷氨酰胺都对大鼠胚胎发育产生不利影响。通过增加 KSOM-R 的体积,大鼠受精卵能够在没有每日培养基更新的情况下以与每天更换培养基的较小体积的受精卵相似的速率和发育能力发育。这些结果代表了对大鼠胚胎培养方法的重要改进,并将有助于更有效地生产大鼠模型。
更新日期:2020-07-01
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