Effect of protein supplementation in potassium simplex optimization medium on preimplantation development of bovine non-transgenic and transgenic cloned embryos
Effect of protein supplementation in potassium simplex optimization medium on preimplantation development of bovine non-transgenic and transgenic cloned embryos
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DOI：
10.1016/j.theriogenology.2004.02.006
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发表时间：
2004-11-01
期刊：
影响因子：
2.8
通讯作者：
Hwang, WS
中科院分区：
文献类型：
--
作者：
Bhuiyan, MMU;Cho, JK;Hwang, WS
本研究探讨了在含钾单纯优化培养液（KSOM）中添加蛋白质对牛附植前胚胎发育的影响。将体外受精（IVF）（实验1）、非转基因（实验2）和转基因克隆胚胎（实验3）在添加0.8%BSA（KSOM-BSA）、10%FBS（KSOM-FBS）或0.01%PVA（KSOM-PVA）的KSOM中培养192 In。用人α 1-抗胰蛋白酶基因表达质粒和绿色荧光蛋白（GFP）标记物转染卵丘细胞以产生转基因克隆胚胎。使用补充有0.8%BSA（mSOF-BSA）的改良合成输卵管液（mSOF）作为对照培养基。在实验1中，KSOM-FBS培养的IVF胚胎的卵裂率（69.1%）显著低于KSOM-BSA培养的卵裂率（80.3%）（P < 0.05）。KSOM-PVA培养的胚胎孵化率（2.2%）显著低于KSOM-FBS培养的胚胎（10.8%）（P < 0.05）。在KSOM-FBS中培养的胚泡含有比在mSOF-BSA中培养的胚泡（36.9 +/- 19.2）显著（P < 0.05）更高数量的内细胞团细胞（50.4 +/- 20.2）。在实验2中，KSOM-PVA培养液的囊胚形成率（20.5%）显著低于其他培养液（33.3-38.5%）（P < 0.05）。KSOM-PVA和KSOM-FBS的囊胚孵化率（13.9%和17.1%）显著低于KSOM-BSA和mSOF-BSA（30.8%和33.9%）（P < 0.05）。在KSOM-PVA中培养的囊胚中，总细胞和滋养外胚层细胞的数量（分别为104.6 +/- 32.2和71.7 +/- 25.5）显著（P < 0.05）低于KSOM-BSA（分别为125.7 +/- 39.7和91.7 +/- 36.2）。在实验3中，在胚胎发育、GFP表达和囊胚细胞数方面，未观察到培养组之间的显著差异。总之，本研究表明，KSOM和补充BSA的mSOF在支持牛非转基因和转基因克隆胚胎发育方面同样有效。此外，在牛非转基因克隆胚胎和转基因克隆胚胎之间观察到了对KSOM蛋白补充反应的不同发育能力。(c)2004年爱思唯尔公司All rights reserved.
The present study evaluated the effect of protein supplementation in potassium simplex optimization medium (KSOM) on bovine preimplantation embryo development. The in vitro fertilized (IVF) (Experiment 1), non-transgenic (Experiment 2) and transgenic cloned embryos (Experiment 3) were cultured for 192 In in KSOM supplemented with 0.8% BSA (KSOM-BSA), 10% FBS (KSOM-FBS) or 0.01% PVA (KSOM-PVA). Transfected cumulus cells with an expression plasmid for human alpha 1-antitrypsin gene and a green fluorescent protein (GFP) marker were used to produce transgenic cloned embryos. Modified synthetic oviductal fluid (mSOF) supplemented with 0.8% BSA (mSOF-BSA) was used as a control medium. In Experiment 1, cleavage rate was significantly (P < 0.05) lower (69.1%) in IVF embryos cultured in KSOM-FBS than in KSOM-BSA (80.3%). The rate of hatching/hatched blastocyst formation was significantly (P < 0.05) lower in embryos cultured in KSOM-PVA than in KSOM-FBS (2.2% versus 10.8%). Blastocysts cultured in KSOM-FBS contained significantly (P < 0.05) higher numbers of inner cell mass cells (50.4 +/- 20.2) than those cultured in mSOF-BSA (36.9 +/- 19.2). In Experiment 2, the rate of blastocyst formation was significantly (P < 0.05) lower (20.5%) in embryos cultured in KSOM-PVA than in other culture media (33.3-38.5%). The rate of hatching/hatched blastocysts was significantly (P < 0.05) lower in KSOM-PVA (13.9%) and KSOM-FBS (17.1%) than in KSOM-BSA (30.8%) and mSOF-BSA (33.9%). The numbers of total and trophectoderm cells (104.6 +/- 32.2 and 71.7 +/- 25.5, respectively) were significantly (P < 0.05) lower in blastocysts cultured in KSOM-PVA than in KSOM-BSA (125.7 +/- 39.7 and 91.7 +/- 36.2, respectively). In Experiment 3, no significant differences in embryo development, GFP expression and blastocyst cell numbers were observed among the culture groups. In conclusion, the present study demonstrated that KSOM and mSOF supplemented with BSA were equally effective in supporting development of bovine non-transgenic and transgenic cloned embryos. Moreover, different developmental competence in response to protein supplementation of KSOM was observed between bovine non-transgenic and transgenic cloned embryos. (c) 2004 Elsevier Inc. All rights reserved.