摘要
【目的】建立高效的猪体细胞克隆胚胎的体外培养体系。【方法】比较了克隆胚胎体外培养过程中高、低氧分压(20%O2和7%O2)和2种培养基(NCSU-23和PZM-3)及添加表皮生长因子(EGF)和碱性成纤维促进因子(bFGF)对胚胎的囊胚发育率及囊胚细胞数的影响。【结果】(1)与高氧(20%O2)气相相比,低氧(7%O2)条件下NCSU-23显著提高了胚胎的囊胚率和细胞数(P<0.05,13.7%vs.8.2%;46.3vs.31.0),培养基PZM-3只能显著提高囊胚细胞数(P<0.05,49.4vs.36.6);(2)与NCSU-23相比,PZM-3无论在高氧还是低氧条件,囊胚率和囊胚细胞数都较高;(3)NCSU-23中添加10ng·ml-1EGF或bFGF对胚胎的囊胚发育率(P<0.05,25.5%,31.7%vs.15.1%)有显著提高,但不能提高囊胚细胞数。【结论】低氧条件有利于胚胎的早期发育;PZM-3无论在高氧还是低氧条件下,都是胚胎发育的一个效果稳定、高效的培养基;胚胎培养基中添加EGF或bFGF能提高胚胎的囊胚发育率。
[Objective] This research focused on establishing a more efficient in vitro culture system for porcine somatic cell nuclear transfer （SCNT） embryos. [ Method ] Systematical study had been carded on effect of oxygen tension （20% O2 and 7% O2）, culture media （NCSU-23 and PZM-3） and growth factors （EGF, bFGF） on in vitro development of porcine SCNT embryos, including blastocyst formation rate and cell number per blastocyst. [Result] In experiment 1, Compared with 20% O2, NCSU-23 in low oxygen （7% O2） greatly improved the blastocyst rate and cell number of cloned embryos （P〈0.05, 13.7% vs. 8.2%; 46.3 vs. 31.0） .Cloned embryos in PZM-3 and 7 % O2 only had a greater cell number （P〈0.05, 49.4 vs. 36.6）. Compared with NCSU-23, PZM-3 supported a higher blastocyst rate, regardless of atmosphere. In experilnent 2, 2-4 cell stage cloned embryos cultured in NCSU-23 supplemented with 10 ng.ml1 EGF or bFGF had a significantly greater developmental ability to blastocyst than the control （P〈0.05, 255%, 31.7% vs. 15.1%）. [Conclusion] Low oxygen atmosphere improved blastocyst rate and total cell number; Regardless of atmosphere, PZM-3 supported a higher development rate and quality of porcine SCNT embryos. EGF and bFGF may be necessarily beneficial to the blastocyst formation.
出处
《中国农业科学》
CAS
CSCD
北大核心
2008年第4期1186-1191,共6页
Scientia Agricultura Sinica
基金
国家"十五"科技攻关项目(2004BA717B)
关键词
猪
体细胞核移植
氧分压
生长因子
培养基
Pig
Somatic cell nuclear transfer
Oxygen tension
Growth factor
Culture medium
