巨噬细胞集落刺激因子(GM-CSF)对小鼠胚胎体外发育的影响  

Effect of granulocyte-macrophage colony-stimulating factor on in vitro development of mouse embryos

作　　者：周娜汝[1,2] 张宇[1,2] 吴蓉花[1,2] 李运生[1,2] 张运海[1,2] 

机构地区：[1]安徽农业大学动物科技学院,合肥230036 [2]安徽地方畜禽遗传资源保护与生物育种省级实验室,合肥230036

出　　处：《安徽农业大学学报》2013年第1期83-89,共7页Journal of Anhui Agricultural University

基　　金：国家高技术研究发展计划(863)项目(2011AA100307-04)资助

摘　　要：以小鼠自然受精与体外受精胚胎为模型,探索鼠源巨噬细胞集落刺激因子(GM-CSF)在小鼠胚胎早期发育过程中的生理功能。分别使用添加不同剂量(对照组:0 ng.mL-1;试验组:0.5、2和10 ng.mL-1)GM-CSF的化学限定培养基连续培养小鼠自然受精、体外受精原核期与2细胞期胚胎,检测其胚胎着床前发育效率(卵裂率、囊胚率)及囊胚的质量(囊胚总细胞数、ICM/总细胞数的比率、凋亡指数)。结果发现,对自然受精胚胎而言,原核时期添加不同剂量的GM-CSF,其卵裂率、囊胚总细胞数、ICM/总细胞数的比率在各组之间均差异不显著,但10 ng.mL-1试验组的囊胚率显著低于对照组(61.6±5.1)%(P<0.05);2细胞时期添加不同剂量的GM-CSF,其囊胚率、囊胚总细胞数、ICM/总细胞数的比率在各组之间均差异不显著,但试验组中的囊胚凋亡指数均显著低于对照组(P<0.05)),并且试验组中2 ng.mL-1的囊胚凋亡指数最低,显著低于0.5 ng.mL-1(P<0.05),其他试验组之间的囊胚凋亡指数没有统计学上的差异性。对体外受精胚胎来说,原核时期添加GM-CSF,其卵裂率、囊胚率、囊胚总细胞数、ICM/总细胞数的比率在各组之间均无显著性差异;在2细胞时期,10 ng.mL-1组的囊胚率显著低于对照组及其他2个试验组(P<0.05),但各组之间的囊胚总细胞数、ICM/总细胞数的比率均无显著性差异。本研究结果表明,在化学限定培养基中添加一定剂量的GM-CSF有利于改善小鼠自然受精与体外受精囊胚的质量,但剂量过高可能不利于小鼠受精胚胎的早期发育。The aim of this study was to explore the physiological function of granulocyte-macrophage col- ony-stimulating factor （GM-CSF） during early development of mouse embryos derived from in vivo natural mat- ing （IVN） or in vitro fertilization （IVF）. Chemically defined medium （CDM） with different concentrations of GM-CSF （the control group： 0 ng·mL^-1; the treatment groups： 0.5, 2 and 10ng·mL^-1） was used for embryo culture, and the cleavage rate, formation rate and quality of blastocysts （total cell number, inner cell mass （ICM）/total cell number, the index of apoptosis） of the embryos were examined. Regarding to the cleavage rate, the total cell number and the ICM/total cell number, no significant difference was found when GM-CSF added from the pronu- clear stage IVN embryos, but the blastocyst rate of 10 ng.mL1 group was significantly lower than that of the con- trol group （40.3%4-6.7% vs. 61.6%4-5.1%; P〈0.05）. For 2-cell stage IVF embryos, in terms of the blastocyst rate, the total cell number, the ICM/total cell number, no significant difference existed among groups, but the index of apoptosis in blastocyst from three treatment groups was obviously decreased comPared to the control group （P〈0.05）. Moreover, the index of apoptosis of blastocyst from 2 ng-mL-1 group was the lowest, even lower than that from 0.5 ng/mL group （P〈0.05）. Apart from that, other treatment groups had no statistical difference, in the blastocyst apoptosis index. As for embryos in the pronuclear stage derived from IVF, no significant difference was observed among groups, regarding the clevage rate, the blastocyst rate, the total cell number and the ICM/total cell number. When GM-CSF was added just from 2-cell stage IVF embryos, the blastocyst rate in the 10 ng.mL-1 group was significantly lower than that in the control group and the other two test groups （P〈 0.05）, but the total cell number in blastocysts, the ICM/total cell number in all groups were not significant different. In conc

关 键 词：GM-CSF 化学限定培养基 小鼠 早期胚胎 体外培养 

分 类 号：S814.8[农业科学—畜牧学] S865.13[农业科学—畜牧兽医]