CAJ | 학술논문

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.
이소서자연수정여체외수정배태위모형,탐색서원거서세포집락자격인자(GM-CSF)재소서배태조기발육과정중적생리공능.분별사용첨가불동제량(대조조:0 ng·mL-1;시험조:0.5、2화10 ng·mL-1)GM-CSF적화학한정배양기련속배양소서자연수정、체외수정원핵기여2세포기배태,검측기배태착상전발육효솔(란렬솔、낭배솔)급낭배적질량(낭배총세포수、ICM/총세포수적비솔、조망지수).결과발현,대자연수정배태이언,원핵시기첨가불동제량적 GM-CSF,기란렬솔、낭배총세포수、ICM/총세포수적비솔재각조지간균차이불현저,단10 ng·mL-1시험조적낭배솔현저저우대조조(61.6±5.1)%(P <0.05);2세포시기첨가불동제량적GM-CSF,기낭배솔、낭배총세포수、ICM/총세포수적비솔재각조지간균차이불현저,단시험조중적낭배조망지수균현저저우대조조(P<0.05)),병차시험조중2 ng·mL-1적낭배조망지수최저,현저저우0.5 ng·mL-1(P<0.05),기타시험조지간적낭배조망지수몰유통계학상적차이성.대체외수정배태래설,원핵시기첨가GM-CSF,기란렬솔、낭배솔、낭배총세포수、ICM/총세포수적비솔재각조지간균무현저성차이;재2세포시기,10 ng·mL-1조적낭배솔현저저우대조조급기타2개시험조(P<0.05),단각조지간적낭배총세포수、ICM/총세포수적비솔균무현저성차이.본연구결과표명,재화학한정배양기중첨가일정제량적 GM-CSF 유리우개선소서자연수정여체외수정낭배적질량,단제량과고가능불리우소서수정배태적조기발육.
The aim of this study was to explore the physiological function of granulocyte-macrophage colo-ny-stimulating factor (GM-CSF) during early development of mouse embryos derived from in vivo natural mating (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 10 ng·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 pro-nuclear stage IVN embryos, but the blastocyst rate of 10 ng·mL-1 group was significantly lower than that of the control group (40.3%±6.7%vs. 61.6%±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 in-dex 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 conclusion, CDM within certain dose of GM-CSF might be helpful to improve the quality of blastocyst derived from either in vivo fertilization or in vitro fertilization in mouse embryos, but over-high concentration of GM-CSF might be harmful to the early development of mouse embryos.