Jiangsu Key Laboratory for Molecular and Medical Biotechnology, College of Life Sciences, Nanjing Normal University, Nanjing, 210023, PR China.
Department of Biology, Central Connecticut State University, New Britain, CT, 06050, USA.
Theriogenology. 2019 Dec;140:109-116. doi: 10.1016/j.theriogenology.2019.08.015. Epub 2019 Aug 13.
The study was designed to determine the impact of magnesium (Mg) on bovine embryo development. We found that two commercially available sources of bovine serum albumin (BSA) and fetal bovine serum (FBS) contained different amounts of Mg residue: 4 ppm in ICPbio BSA, 114 ppm in Sigma BSA, and 44 ppm in FBS. When CR1 was used as basal medium, PVA and ICPbio BSA produced the lowest blastocyst yield (2.2-2.3%), whereas Sigma BSA increased blastocyst yield to 18.9% (P < 0.05). Supplementation of 1.4 mM MgCl into the medium increased the blastocyst rate in the ICPbio BSA group (29.4%) but not in the PVA group (5.4%; P < 0.05) to a level comparable to that of the FBS group (33.7%; P > 0.05). We next found that increasing concentrations of MgCl in the culture medium (ICPbio BSA) elevated blastocyst rate from 2.6% (0 mM), 38.4% (0.35 mM) to 50.2% (1.4 mM; P < 0.05), further maintained at 44.9% (2.1 mM) and 43.4% (2.8 mM) (P > 0.05). However, blastocyst rate was reduced to 31.4% (4.2 mM) and 29.4% (5.6 mM) when MgCl supplement was increased (P < 0.05). Comparable blastocyst development was achieved in both ICPbio BSA (30.0-33.1%) and Sigma BSA (37.4-38.7%) groups when 1.4 mM Mg was supplemented regardless of its source (MgCl vs. MgSO; P > 0.05). In embryo transfer experiments, higher rates of pregnancy (54.3 vs. 41.5%) and calving (44.3 vs. 32.5%) were achieved in the CR1-Mg-supplemented BSA group compared with the FBS group with co-culture, respectively (P < 0.05). These results demonstrate that Mg is a key ion that promotes competent blastocyst and term development. Therefore, a simple and efficient defined medium (CR1-Mg-BSA) can successfully replace complex serum and somatic cell co-culture.
本研究旨在探究镁(Mg)对牛胚胎发育的影响。我们发现,两种市售牛血清白蛋白(BSA)和胎牛血清(FBS)来源的产品含有不同量的镁残留:ICPbio BSA 中含有 4ppm 的镁残留,Sigma BSA 中含有 114ppm 的镁残留,FBS 中含有 44ppm 的镁残留。当 CR1 被用作基础培养基时,PVA 和 ICPbio BSA 产生的囊胚产量最低(2.2-2.3%),而 Sigma BSA 则将囊胚产量提高到 18.9%(P<0.05)。在培养基中补充 1.4mM MgCl2 可提高 ICPbio BSA 组的囊胚率(29.4%),但对 PVA 组(5.4%;P<0.05)没有影响,两组囊胚率与 FBS 组(33.7%;P>0.05)相当。接下来,我们发现,随着培养基中 MgCl2 浓度的升高(ICPbio BSA),囊胚率从 2.6%(0mM)、38.4%(0.35mM)提高到 50.2%(1.4mM;P<0.05),进一步保持在 44.9%(2.1mM)和 43.4%(2.8mM)(P>0.05)。然而,当补充的 MgCl2 增加时(4.2mM 和 5.6mM),囊胚率分别降低至 31.4%和 29.4%(P<0.05)。无论镁的来源(MgCl2 与 MgSO4)如何,在补充 1.4mM Mg 后,ICPbio BSA(30.0-33.1%)和 Sigma BSA(37.4-38.7%)组的囊胚发育情况相当(P>0.05)。在胚胎移植实验中,与 FBS 组共培养相比,CR1-Mg 补充的 BSA 组的妊娠率(54.3% vs. 41.5%)和产犊率(44.3% vs. 32.5%)均更高(P<0.05)。这些结果表明,镁是促进囊胚和足月发育的关键离子。因此,简单有效的定义培养基(CR1-Mg-BSA)可以成功替代复杂的血清和体细胞共培养。