Tsuji Y, Iwasaki T, Ogata H, Matsumoto Y, Kokeguchi S, Matsumura K, Hyon S H, Shiotani M
Hanabusa Women's Clinic, Kobe city, Hyogo, Japan.
School of Materials Science, Japan Advanced Institute of Science and Technology, Nomi, Ishikawa, Japan.
Cryo Letters. 2017 Jan/Feb;38(1):1-6.
In the vitrification of embryos, dimethyl sulfoxide (DMSO) is one of the most effective cryoprotectant agents (CPAs), but cytotoxic effects of DMSO on embryos are well known. Carboxylated poly-L-lysine (CPLL) has been identified as an effective cryoprotectant of cultured cell lines and mammalian oocytes.
To evaluate the efficacy and safety of CPLL as a CPA for developmental stage embryos.
Mouse 8-cell embryos and blastocysts were vitrified with ethylene glycol (EG), DMSO/EG, or CPLL/EG and the developmental potency assessed in vitro.
In 8-cell embryos, there were no differences between the levels of survival and developmental progress into the blastocyst stage in each solution. At the blastocyst stage, the proportion of dead cells was significantly higher in the EG compared with other solutions. In contrast, there were no differences between the DMSO/EG and CPLL/EG.
These results indicate that CPLL can be used as a replacement for DMSO in the vitrification of mouse embryos.
在胚胎玻璃化冷冻中,二甲基亚砜(DMSO)是最有效的冷冻保护剂(CPA)之一,但DMSO对胚胎的细胞毒性作用是众所周知的。羧化聚-L-赖氨酸(CPLL)已被确定为培养细胞系和哺乳动物卵母细胞的有效冷冻保护剂。
评估CPLL作为发育阶段胚胎CPA的有效性和安全性。
将小鼠8细胞胚胎和囊胚用乙二醇(EG)、DMSO/EG或CPLL/EG进行玻璃化冷冻,并在体外评估其发育能力。
在8细胞胚胎中,每种溶液的存活率和发育至囊胚阶段的进展水平之间没有差异。在囊胚阶段,与其他溶液相比,EG中死细胞的比例显著更高。相比之下,DMSO/EG和CPLL/EG之间没有差异。
这些结果表明,CPLL可在小鼠胚胎玻璃化冷冻中用作DMSO的替代品。