Karagenc Levent, Lane Michelle, Gardner David K
Bahceci Women Health Care Centre and German Hospital, Istanbul, Turkey.
Reprod Biomed Online. 2005 Apr;10(4):511-8. doi: 10.1016/s1472-6483(10)60829-2.
The aim of the current study was to examine the effects of granulocyte-macrophage colony-stimulating factor (GM-CSF) on the development and differentiation of preimplantation mouse embryos from different strains and under different culture conditions. Embryos from F1 hybrid mice were cultured in a modified G1 medium lacking amino acids and EDTA (simple G1), human tubal fluid medium (HTF) or in G1/G2 sequential media, supplemented with GM-CSF (0, 2, 4, 8, and 16 ng/ml). Embryos from CF1 mice were subsequently cultured in G1/G2 with (5 mg/ml) or without HSA, in the absence or presence of GM-CSF (2 ng/ml). GM-CSF had no effect at any concentration on F1 embryo development and blastocyst cell numbers, irrespective of the culture media used. Similarly, GM-CSF had no effect on CF1 blastocyst development. However, a stimulatory effect of GM-CSF was evident on total blastocyst cell number and ICM development when CF1 embryos were cultured in the absence of HSA. When HSA was present in the media the beneficial effect of GM-CSF was negated. There was no difference in the number of apoptotic cells in CF1 blastocysts when G1/G2 were supplemented with GM-CSF with or without HSA. These data indicate that there is no beneficial effect of supplementing either simple (simple G1 or HTF) or more complete (G1/G2) media with GM-CSF when protein is present in the medium. However, when culture conditions are suboptimal and non-physiological, i.e. the absence of protein, GM-CSF stimulates development of both total cell numbers and ICM development of CF1 blastocysts.
本研究的目的是检测粒细胞-巨噬细胞集落刺激因子(GM-CSF)对不同品系小鼠植入前胚胎在不同培养条件下发育和分化的影响。将F1杂交小鼠的胚胎培养于缺乏氨基酸和乙二胺四乙酸(EDTA)的改良G1培养基(简易G1)、人输卵管液培养基(HTF)或添加了GM-CSF(0、2、4、8和16 ng/ml)的G1/G2序贯培养基中。随后,将CF1小鼠的胚胎在添加或不添加人血清白蛋白(HSA,5 mg/ml)的情况下,于有无GM-CSF(2 ng/ml)存在时培养于G1/G2中。无论使用何种培养基,GM-CSF在任何浓度下对F1胚胎发育和囊胚细胞数量均无影响。同样,GM-CSF对CF1囊胚发育也无影响。然而,当CF1胚胎在无HSA的情况下培养时,GM-CSF对囊胚总细胞数量和内细胞团(ICM)发育具有明显的刺激作用。当培养基中存在HSA时,GM-CSF的有益作用被抵消。在G1/G2中添加或不添加HSA的情况下,CF1囊胚中凋亡细胞的数量没有差异。这些数据表明,当培养基中存在蛋白质时,在简易培养基(简易G1或HTF)或更完全的培养基(G1/G2)中添加GM-CSF均无有益作用。然而,当培养条件不理想且不符合生理状态时,即缺乏蛋白质时,GM-CSF可刺激CF1囊胚的总细胞数量和ICM发育。