Viability and developmental potential of porcine blastocysts preserved for short term in a chemically defined medium at ambient temperature
Viability and developmental potential of porcine blastocysts preserved for short term in a chemically defined medium at ambient temperature
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环境温度下化学成分确定的培养基中短期保存的猪囊胚的活力和发育潜力
DOI：
10.1111/rda.14095
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发表时间：
2022
影响因子：
1.7
通讯作者：
Otoi Takeshige
中科院分区：
文献类型：
--
作者：
Lin Qingyi;Le Quynh Anh;Takebayashi Koki;Hirata Maki;Tanihara Fuminori;Thongkittidilok Chommanart;Sawamoto Osamu;Kikuchi Takeshi;Otoi Takeshige
本研究开发了一种在环境温度下液体储存体外猪囊胚24 h的有效方法。我们评价了新的化学成分确定的培养基（细胞清洗和保存液，Cellstor®-W [Cell-W]和细胞悬液和保存液，Cellstor®-S [Cell-S]）对短期储存的影响。在第一个实验中，将体外衍生的囊胚在Cell-W溶液、Cell-S溶液和猪胚胎培养（PBM）培养基中于25 ℃下储存24小时。Cell-S和Cell-W溶液之间储存的囊胚的存活率和发育率没有差异，但在Cell-S溶液中储存后存活的胚胎的总细胞数显著高于Cell-W溶液中的总细胞数（p<0.05）。在第二个实验中，使用Cell-S溶液在20°C、25°C和30°C下储存体外衍生的囊胚。与在25°C或30°C下储存相比，在20°C下储存导致储存的胚泡的存活率和发育率显著降低。在25°C和30°C下储存之间没有观察到存活率和发育率的差异，但在25°C下储存和培养后对胚胎质量的损害显著低于30°C。在第三个实验中，Cell-S溶液补充有β-巯基乙醇和姜黄素，单独或组合作为抗氧化剂。虽然补充姜黄素并没有提高存活率，但与不含抗氧化剂的对照囊胚相比，它显著增加了储存囊胚的发育率。总之，当猪囊胚在25°C下储存24小时时，Cell-S溶液可有效维持体外胚胎的存活和发育。
This study developed an efficient method for liquid storage of in vitro‐derived porcine blastocysts at ambient temperature for 24 hr. We evaluated the effects of new chemically defined media (cell wash and preservation solution, Cellstor®‐W [Cell‐W] and cell suspension and preservation solution, Cellstor®‐S [Cell‐S]) for short‐term storage. In the first experiment, in vitro‐derived blastocyst were stored at 25ºC for 24 hr in Cell‐W solution, Cell‐S solution and pig embryo culture (PBM) medium. There were no differences in the rates of survival and development of stored blastocysts between the Cell‐S and Cell‐W solutions, but the total cell number of embryos that survived after storage in Cell‐S solution was significantly higher (p< .05) than that in the Cell‐W solution. In the second experiment, Cell‐S solution was used to store the in vitro‐derived blastocysts at 20°C, 25°C and 30°C. Storage at 20°C resulted in a significant decrease in the rates of survival and development of stored blastocysts compared to storage at 25°C or 30°C. No differences in survival and development rates were observed between storage at 25°C and 30°C, but the damage to the embryo quality after storage and culture was significantly lower at 25°C than at 30°C. In the third experiment, Cell‐S solution was supplemented with β‐mercaptoethanol and curcumin, either alone or in combination, as antioxidant agents. Although the supplementation with curcumin did not improve survival, it significantly increased the development rate of stored blastocysts compared with the control blastocysts stored without antioxidants. In conclusion, when porcine blastocysts were stored at 25°C for 24 hr, a Cell‐S solution may be effective for maintaining the survival and development of in vitro embryos.