Parthenogenetic activation of rabbit oocytes and effects of different additives on development of activated embryos
10.3760/cma.j.issn.1671-8925.2013.01.007
- VernacularTitle:兔卵母细胞孤雌激活及不同添加物对激活胚胎发育的影响
- Author:
Yu-Xi ZOU
1
;
Jiang-Hua SHANG
;
Lin-Sha QIN
;
Yan-Ping TANG
;
Xiao-Dan JIANG
Author Information
1. 510282广州,南方医科大学珠江医院神经外科,广东神经外科研究所,广东省脑功能修复与再生重点实验室
- Keywords:
Embryonic development;
Parthenogenetic activation;
Oocyte
- From:
Chinese Journal of Neuromedicine
2013;12(1):29-33
- CountryChina
- Language:Chinese
-
Abstract:
Objective To investigate an appropriate activation method and culture system in vitro of rabbit parthenogenetic embryos to make an experimental foundation for therapeutic cloning.Methods Mature rabbit oocytes with polar body 1 (PB 1) were divided randomly into four groups and accepted,respectively,parthenogenetic activation with 1.2 kV/cm,1.6 kV/cm and 2.0 kV/cm electricity stimulation or 5 μg/mL ionomycin (ION) chemical treatment; the effects of these different treatments on activation efficiency were compared.The activated oocytes were cultured,respectively,in M199 supplemented with various concentrations of leukaemia inhibitory factor (LIF,0,10,20,40 and 80 ng/mL) or insulin+transferrin+sodium selenite (ITS,0×,1 × and 2×); and then,the rate of cleaved oocytes,percentage of morulas/blastocysts and total number of blastocysts were compared between each two groups.Results (1) Different treatments on the oocytes showed significantly different activation effects.The survival rate of oocytes activated in 5 μg/mL ION for 4 min were significantly higher than that of ones stimulated by 2.0 kV/cm electricity treatment (P=0.016).The percentages of cleaved oocytes,developing 8-16 cells and morulas/blastocysts in the ION treatment group were also significantly higher as compared with those in the 1.2 kV/cm electric treatment group (P=0.000,P=0.002 andP=0.026,respectively).(2) Significant difference in the percentage of morulas/blastocysts was noted between 20 ng/mL and 80 ng/mL of LIF supplementation (P=0.003); in addition,the total number ofblastocysts in the medium supplemented with 20 ng/mL LIF was significantly different to those with 40 ng/mL and 80 ng/mL,respectively (P=0.011,P=0.002); In experiment of ITS supplementation,significant difference in the percentage ofmorulas/blastocysts was shown between 1 × and 2× ITS supplementation (P=0.003); in addition,the total number of blastocysts in the medium supplemented with 0× or 1 × ITS (166 and 147 cells) was significantly different as compared with that with 2× ITS (78 cells,P=0.015,P=0.044).Conclusion The parthenogenesis activation of the New Zealand rabbit's oocytes treated with ION+6-DMAP shows a higher rate of cleaved oocytes and blastocysts than that with electricity stimulation; the subsequent development of parthenogenetic embryos in vitro could be significantly improved by supplementing with 20 ng/mL LIF and 1× ITS.
