Kelley Rebecca L, Gardner David K
School of BioSciences, The University of Melbourne, Parkville, Victoria 3010, Australia.
School of BioSciences, The University of Melbourne, Parkville, Victoria 3010, Australia.
Reprod Biomed Online. 2017 May;34(5):441-454. doi: 10.1016/j.rbmo.2017.02.001. Epub 2017 Feb 15.
Single embryo culture is suboptimal compared with group culture, but necessary for embryo monitoring, and culture systems should be improved for single embryos. Pronucleate mouse embryos were used to assess the effect of culture conditions on single embryo development. Single culture either before or after compaction reduced cell numbers (112.2 ± 3.1; 110.2 ± 3.5) compared with group culture throughout (127.0 ± 3.4; P < 0.05). Reduction of media volume from 20 µl to 2 µl increased blastocyst cell numbers in single embryos cultured in 5% oxygen (84.4 ± 3.2 versus 97.8 ± 2.8; P < 0.05), but not in 20% oxygen (55.2 ± 2.9 versus 57.1 ± 2.8). Culture in microwell plates for the EmbryoScope and Primo Vision time-lapse systems changed cleavage timings and increased inner cell mass cell number (24.1 ± 1.0; 23.4 ± 1.2) compared with a 2 µl microdrop (18.4 ± 1.0; P < 0.05). Addition of embryo-conditioned media to single embryos increased hatching rate and blastocyst cell number (91.5 ± 4.7 versus 113.1 ± 4.4; P < 0.01). Single culture before or after compaction is therefore detrimental; oxygen, media volume and microwells influence single embryo development; and embryo-conditioned media may substitute for group culture.
与群体培养相比，单个胚胎培养并不理想，但对于胚胎监测是必要的，并且应改进单个胚胎的培养系统。使用原核期小鼠胚胎来评估培养条件对单个胚胎发育的影响。与全程群体培养（127.0±3.4）相比，在致密化之前或之后进行单个培养会减少细胞数量（112.2±3.1；110.2±3.5）（P<0.05）。将培养基体积从20微升减少到2微升可增加在5%氧气中培养的单个胚胎的囊胚细胞数量（84.4±3.2对97.8±2.8；P<0.05），但在20%氧气中则不然（55.2±2.9对57.1±2.8）。与2微升微滴培养相比，在用于EmbryoScope和Primo Vision延时系统的微孔板中培养会改变卵裂时间并增加内细胞团细胞数量（24.1±1.0；23.4±1.2）（18.4±1.0；P<0.05）。向单个胚胎添加胚胎条件培养基可提高孵化率和囊胚细胞数量（91.5±4.7对113.1±4.4；P<0.01）。因此，在致密化之前或之后进行单个培养是有害的；氧气、培养基体积和微孔会影响单个胚胎发育；并且胚胎条件培养基可能替代群体培养。
