Turpeenniemi-Hujanen T, Feinberg R F, Kauppila A, Puistola U
Oulu University Central Hospital, Finland.
Fertil Steril. 1995 Jul;64(1):132-8.
To study the role of the extracellular matrix (ECM) proteins in supporting the development and implantation competence of human embryos.
Expression of an implantation site adhesive glycoprotein, oncofetal fibronectin, and basement membrane collagen-degrading matrix metalloproteinase-2 and -9 were studied in cultured human embryos. The ability of exogenously added laminin and fibronectin to enhance hatching and matrix metalloproteinase-2 expression was quantitated also.
Fifty-four women with tubal factor infertility enrolled in the IVF program at the University Hospital of Oulu agreed to participate by providing 20 residual oocytes and 227 residual early embryos for this study.
The presence of oncofetal fibronectin immunoreactive protein was assayed by immunocytochemical staining with two specific monoclonal antibodies, FDC-6 and X18A4. These antibodies bind to specific and distinct epitopes within tumor and trophoblast-derived oncofetal fibronectin. Changes in embryo matrix metalloproteinase-2 production were measured by zymography and confirmed by immunocytochemical staining.
Intracellular oncofetal fibronectin was identified within blastomeres of early stage embryos. The immunoreactivity of oncofetal fibronectin in the zona pellucida was associated with fragmentation and dissolution of the zona. Exogenously added laminin or adult-type fibronectin significantly increased the hatching rate of the cultured embryos. Embryos cultured with added adult-type fibronectin or trophoblast-derived oncofetal fibronectin stimulated the matrix metalloproteinase-2 production (72-kd type IV collagenase) by 2.25 +/- 0.16-fold when compared with control embryos (mean +/- SD).
Embryonic production of specific ECM proteins, such as oncofetal fibronectin, appears to be important for the morphological and biochemical development of human preimplantation embryos. Moreover, ECM proteins promote acquisition of the adhesive and degradative properties required by human embryos for successful implantation.
研究细胞外基质（ECM）蛋白在支持人类胚胎发育和着床能力方面的作用。
在培养的人类胚胎中研究着床部位黏附糖蛋白、癌胚纤连蛋白以及基底膜胶原降解基质金属蛋白酶-2和-9的表达。还对外源性添加的层粘连蛋白和纤连蛋白增强孵化和基质金属蛋白酶-2表达的能力进行了定量分析。
奥卢大学医院体外受精项目中54名输卵管因素不孕症女性同意参与，为本研究提供20个剩余卵母细胞和227个剩余早期胚胎。
用两种特异性单克隆抗体FDC-6和X18A4通过免疫细胞化学染色检测癌胚纤连蛋白免疫反应性蛋白的存在。这些抗体与肿瘤和滋养层来源的癌胚纤连蛋白内的特定且不同的表位结合。通过酶谱法测量胚胎基质金属蛋白酶-2产生的变化，并通过免疫细胞化学染色进行确认。
在早期胚胎的卵裂球内鉴定出细胞内癌胚纤连蛋白。透明带中癌胚纤连蛋白的免疫反应性与透明带的碎片化和溶解有关。外源性添加层粘连蛋白或成人型纤连蛋白显著提高了培养胚胎的孵化率。与对照胚胎相比，添加成人型纤连蛋白或滋养层来源的癌胚纤连蛋白培养的胚胎刺激基质金属蛋白酶-2产生（72-kd IV型胶原酶）增加了2.25±0.16倍（平均值±标准差）。
特定ECM蛋白如癌胚纤连蛋白的胚胎产生似乎对人类植入前胚胎的形态和生化发育很重要。此外，ECM蛋白促进人类胚胎成功着床所需的黏附性和降解性的获得。
