Battye K M, Ammit A J, O'Neill C, Evans G
Human Reproduction Unit, Royal North Shore Hospital, St Leonards, NSW, Australia.
J Reprod Fertil. 1991 Nov;93(2):507-14. doi: 10.1530/jrf.0.0930507.
Embryos were collected from superovulated ewes on Day 2 (2-8 cell), Day 4 (8-16 cell) and Day 6 (morula/early blastocyst). Two embryos were cultured in 1 ml of one of four media: (i) Ham's F10 + 4 mg bovine serum albumin (BSA)/ml, (ii) synthetic oviduct fluid medium + 20% human serum, (iii) Quinn's human tubal fluid medium (HTF) + 3 mg BSA/ml or (iv) HTF + 10% acid-treated fetal calf serum for 24 h. They were transferred to fresh media of the same type and their further development was monitored. A quantitative bioassay and radioimmunoassay was used to measure the concentration of platelet-activating factor (PAF, 1-o-alkyl-2-acetyl-sn-glyceryl-3-phosphocholine) produced. Following extraction and partial purification, 21/95 (22.1%) of the embryo-conditioned media samples had PAF concentrations greater than that measured in corresponding control media. This was designated as embryo-derived PAF and the corresponding cultures were termed 'PAF-positive'. PAF was produced by embryos at all three developmental stages examined and in each of the four media used, and the average amount of PAF produced was 60.9 +/- 9.8 pmol/embryo/24 h. However, neither the developmental stage of the embryo, nor the type of media affected the proportion of PAF-positive cultures nor the amount of PAF produced during culture. Thus, it is demonstrated for the first time that early ovine embryos can secrete PAF in vitro, and that there is considerable variability in their capacity for PAF secretion.
在第2天（2-8细胞期）、第4天（8-16细胞期）和第6天（桑葚胚/早期囊胚期）从超排母羊收集胚胎。将两个胚胎培养于1 ml四种培养基之一中：（i）Ham's F10 + 4 mg/ml牛血清白蛋白（BSA），（ii）合成输卵管液培养基 + 20%人血清，（iii）Quinn's人输卵管液培养基（HTF） + 3 mg/ml BSA，或（iv）HTF + 10%酸处理胎牛血清，培养24小时。将它们转移至相同类型的新鲜培养基中，并监测其进一步发育情况。采用定量生物测定和放射免疫测定法测量所产生的血小板活化因子（PAF，1-油酰基-2-乙酰基-sn-甘油-3-磷酸胆碱）浓度。经过提取和部分纯化后，95个胚胎条件培养基样本中有21个（22.1%）的PAF浓度高于相应对照培养基中测得的浓度。这被指定为胚胎源性PAF，相应培养物被称为“PAF阳性”。在所检查的所有三个发育阶段的胚胎以及所用的四种培养基中的每一种中均产生了PAF，PAF的平均产生量为60.9±9.8 pmol/胚胎/24小时。然而，胚胎的发育阶段和培养基类型均未影响PAF阳性培养物的比例或培养期间产生的PAF量。因此，首次证明早期绵羊胚胎能够在体外分泌PAF，并且它们的PAF分泌能力存在相当大的变异性。
