Liu L P, Chan S T, Ho P C, Yeung W S
Department of Zoology, University of Hong Kong, Hong Kong.
Hum Reprod. 1998 Jun;13(6):1613-9. doi: 10.1093/humrep/13.6.1613.
Embryotrophic factors from human oviductal cells were partially purified by liquid chromatographic methods. The conditioned medium from human oviductal cell culture was fractionated successively by concanavalin A (Con-A) affinity chromatography, ion-exchange chromatography and gel filtration. The presence of the embryotrophic activity in the eluates was determined by the stimulatory effects on the development of mouse embryos in vitro. The fraction that did not bind to the lectin Con-A possessed no embryotrophic activity. Ion-exchange chromatography separated the glycoproteins that bound to Con-A into five fractions. Three of them significantly enhanced blastulation as well as conceptus formation. Gel filtration further separated these embryotrophic fractions into five fractions. Three of them with molecular weights of 154 +/- 1, 164 +/- 0.2 and 207 +/- 0.3 kDa significantly stimulated blastulation of mouse embryos. The results of this study demonstrated that several embryotrophic factors with different biochemical properties contributed to the embryotrophic effect of the human oviductal cell/mouse embryo co-culture system.
采用液相色谱法对人输卵管细胞中的胚胎营养因子进行了部分纯化。人输卵管细胞培养的条件培养基先后通过伴刀豆球蛋白A(Con-A)亲和色谱、离子交换色谱和凝胶过滤进行分级分离。通过体外对小鼠胚胎发育的刺激作用来确定洗脱液中胚胎营养活性的存在。未与凝集素Con-A结合的部分不具有胚胎营养活性。离子交换色谱将与Con-A结合的糖蛋白分离成五个部分。其中三个部分显著增强了囊胚形成以及孕体形成。凝胶过滤进一步将这些胚胎营养部分分离成五个部分。其中三个分子量分别为154±1、164±0.2和207±0.3 kDa的部分显著刺激了小鼠胚胎的囊胚形成。本研究结果表明,几种具有不同生化特性的胚胎营养因子对人输卵管细胞/小鼠胚胎共培养系统的胚胎营养作用有贡献。