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대한생식의학회지 제31권 제1호 2010년
생쥐모델을 이용한 동결보존제의 독성조사
송촌 산부인과, 서울보건대학 임상병리과, 을지의과대학교 산부인과
양관철, 강희규,이회창,이향흔,고덕성,양현원,박원일,박은주,김세웅
Toxic Effect of Cryoprotectants on Embryo Development in a Murine Model
Kwan Cheal Yang, Hee-Gyoo Kang, Hoi-Chang Lee, Hyang Heun Lee, Duck-Sung Ko, Hyunwon Yang, Won-Il Park, Eun-Joo Park, S. Samuel Kim
Song-Chon OB/GYN Clinic, Department of Biomedical laboratory science, Seoul health college, Seongnam, Department of Obstetrics & Gynecology, Eulji University School of Medicine, Seoul Korea
Objectives: The aim of this study was to assess toxicities of cryoprotectants. Methods: Toxicities of two cryoprotectants, dimethyl sulfoxide (DMSO) and 1,2-propanediol (PROH), were investigated using a murine embryo model. Female F-1 mice were stimulated with gonadotropin, induced ovulation with hCG and mated. Two cell embryos were collected and cultured after exposure to either DMSO or PROH. Embryo development was evaluated up to the blastocyst stage. Blastocysts were stained with bis-benzimide to evaluate the cell count and with terminal deoxynucleotidyl transferase mediated dUTP nick labeling (TUNEL) to assess apoptosis. Results: The total cell count of blastocysts that were treated with DMSO at the 2-cell stage was significantly lower than that were treated with PROH (75.9¡¾27.0) or the control (99.0¡¾18.3) (p<0.001). On comparison of two cryoprotectant treated groups, the DMSO treated group showed a decreased cell count compared with the PROH treated group (p<0.05). Both DMSO (14.2¡¾1.5) and PROH (11.2¡¾1.4) treated groups showed higher apoptosis rates of cells in the blastocyst compared with the control (6.2¡¾0.9, p<0.0001). In addition, the DMSO treated group showed more apoptotic cells than the PROH treated group (p<0.001). Conclusions: The potential toxicity of cryoprotectants was uncovered by prolonged exposure of murine embryos to either DMSO or PROH at room temperature. When comparing two cryoprotective agents, PROH appeared to be less toxic than DMSO at least in a murine embryo model.
키워드 : Objectives: The aim of this study was to assess toxicities of cryoprotectants.
교신저자 : medssk@attglobal.net
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