Miles Jeremy R, Farin Charlotte E, Rodriguez Karina F, Alexander Joseph E, Farin Peter W
Department of Population Health and Pathobiology, North Carolina State University, Raleigh, 27606, USA.
Biol Reprod. 2005 Oct;73(4):663-71. doi: 10.1095/biolreprod.105.040808. Epub 2005 May 18.
The objective of this study was to determine the effects of undefined and semidefined culture systems for in vitro embryo production on angiogenesis and morphometry of bovine placentas during early gestation. Blastocysts produced in vivo were recovered from superovulated Holstein cows and served as controls. Blastocysts produced in vitro were exposed to either serum-supplemented medium with cumulus cell coculture (in vitro-produced with serum; IVPS) or modified synthetic oviductal fluid medium without serum or coculture (mSOF). Single blastocysts from each production system were transferred into heifers. Fetuses and placentas were recovered on Day 70 of gestation. Cotyledonary tissues were obtained for quantification of vascular endothelial growth factor (VEGF) and peroxisome proliferator-activated receptor-gamma (PPARG) mRNA and protein. Samples of placentomes were prepared for immunocytochemistry and histological analysis. Placentas from the mSOF group were heavier and had the fewest placentomes, least placental fluid, and lowest placental efficiency (fetal weight/placental weight) compared with the in vivo and IVPS groups. There was no effect of embryo culture system on volume densities of fetal villi or maternal endometrium within placentomes. The volume density of fetal pyknotic cells was increased in placentomes in the mSOF group compared with the in vivo and IVPS groups. Placentomes in the mSOF group had decreased densities of blood vessels and decreased levels of VEGF mRNA in cotyledonary tissue. In conclusion, compared with placentas from embryos produced in vivo or in vitro using an undefined culture system, placentas from embryos produced in vitro using a semidefined culture system exhibited a greater degree of aberrant development of the placenta during early gestation.
本研究的目的是确定体外胚胎生产中未定义和半定义培养系统对妊娠早期牛胎盘血管生成和形态测量的影响。从超排的荷斯坦奶牛体内回收体内产生的囊胚作为对照。体外产生的囊胚分别暴露于添加血清并与卵丘细胞共培养的培养基(体外血清培养;IVPS)或无血清或共培养的改良合成输卵管液培养基(mSOF)。将每个生产系统的单个囊胚移植到小母牛体内。在妊娠第70天回收胎儿和胎盘。获取子叶组织以定量血管内皮生长因子(VEGF)和过氧化物酶体增殖物激活受体γ(PPARG)的mRNA和蛋白质。制备胎盘小叶样本用于免疫细胞化学和组织学分析。与体内和IVPS组相比,mSOF组的胎盘更重,胎盘小叶最少,胎盘液最少,胎盘效率(胎儿体重/胎盘体重)最低。胚胎培养系统对胎盘小叶内胎儿绒毛或母体子宫内膜的体积密度没有影响。与体内和IVPS组相比,mSOF组胎盘小叶中胎儿固缩细胞的体积密度增加。mSOF组胎盘小叶的血管密度降低,子叶组织中VEGF mRNA水平降低。总之,与使用未定义培养系统在体内或体外产生的胚胎的胎盘相比,使用半定义培养系统在体外产生的胚胎的胎盘在妊娠早期表现出更大程度的胎盘异常发育。