摘要
为探讨不同细胞因子对水牛原始生殖细胞（PGCs）传代培养的影响，将PGCs分别采用A、B、C、D、E、F和G共7种培养基进行培养，即A组：基础液＋10ng／mL白血病抑制因子（LIF）＋10ng／mL碱性成纤维细胞生长因子（bFGF）；B组：基础液＋20ng／mL LIF＋20ng／mL bFGF；C组：基础液＋20ng／mL LIF＋10ng／mL bFGF；D组：基础液＋20ng／mL LIF；E组：基础液＋20ng／mL LIF＋20ng／mL bFGF＋20ng／mL干细胞因子（SCF）；F组：基础液＋20ng／mL LIF＋40ng／mL bFGF＋40ng／mL SCF；G组（对照组）：基础液。将机械法分离的PGCs小集落接种到水牛胎儿成纤维细胞（BEF）饲养层上进行传代培养。结果，原代时，A、B、C、D、E和F组的克隆数目都显著高于对照组（P％0．05），其中E和F组的克隆数目显著高于A组（P〈0．05），而与B、C、D组差异均不显著（P〉0．05）；1～8代时，B、C、D、E、F组的克隆数目显著高于A组（P〈0．05）；对照组传代数仅为2代，A组为5代，而B、C、D、E和F组均为8代以上。结果表明，在传代过程中，LIF起主要作用，20ng／mL的LIF浓度可以满足水牛PGCs传代培养的需要。
To explore the influence of different cytokines on the subculture of buffalo PGCs, PGCs were cultured with media A（basic culture medium＋ 10 ng/mL LIF＋ 10 ng/mL bFGF）, B（basic culture medium＋20 ng/mL LIF＋20 ng/mL bFGF） ,C（basic culture medium＋20 ng/mL LIF＋ 10 ng/mL bFGF） ,D （basic culture medium＋20 ng/mL LIF）,E（basic culture medium＋20, ng/mL LIF＋20 ng/mL bFGF＋20 ng/mL SCF） ,F（basic culture medium＋20 ng/mL LIF＋ 40 ng/mL bFGF＋40 ng/mL SCF） and G（basic culture medium,as control）, respectively. Then PGCs were dissociated by mechanical method and plated into BEF feeder for subculture. Groups A,B,C,D,E and F were significantly higher than group G in the number of the primary PGCs clones（P（0.05）, and groups E and F were significantly higher than group A in the number of the primary PGCs clones （P〈0. 05） ,and no significant difference was observed among groups B,C and D （P〉0.05）. In 1-8 passage subculture, groups B,C,D and F were significantly higher than group A in the number of the PGCs clones （P〈0.05）. In the maximal subculture, groups B,C,D,E and F were at least eight passages, group A was five passages, but group G was only two passages. The results show that the main factor affecting buffalo PGCs subculture is LIF,and 20 ng/mL LIF can meet the need of buffalo PGCs subculture.
出处
《中国兽医科学》
CAS
CSCD
北大核心
2007年第4期350-353,共4页
Chinese Veterinary Science
基金
国家高技术研究发展计划(863)项目(2002AA206651)
广西研究生教育创新计划项目(2006105930905D07)
