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[体外植入模型]

[Implantation model in vitro].

作者信息

Negami A I

机构信息

Department of Obstetrics and Gynecology, Fukui Medical School.

出版信息

Nihon Sanka Fujinka Gakkai Zasshi. 1992 Aug;44(8):949-59.

PMID:1402227
Abstract

Implantation is thought to be interactive and synchronized process between mother and embryo, however, the mechanisms of the early implantation process remain unelucidated. Therefore, the effectiveness of in vitro fertilization and embryo transfer is still poor. As a result; immunohistochemically, type IV collagen and other extracellular matrix components were detected mainly below the epithelial layer. Type I and III collagens were detected diffusely in the stromal layer. In the lower stromal layer and superficial myometrial layer, type V collagen was detected. Human endometrial epithelial cells performed the re-epithelialization following glandular formation. Rabbit endometrial cells performed also re-epithelialization following the fold formation. The stromal cells were invaded into the inner layer of the folding. Estrogen added to the culture media stimulated the glandular formation. Progesterone administration after estrogen priming did not affect the glandular formation, however, the proliferation of the superficial epithelium and the re-epithelialized area were increased. Rabbit blastocysts successfully attached and implanted into the reconstructed endometrium. The development of the implanted embryos was morphologically normal. Human cultured trophoblast cells attached, invaded and penetrated into the extracellular matrix components. On the other hand, using type V collagen coated dishes, trophoblast cells could invade the stromal layer, however, type V collagen layer did not permit the trophoblast cells to invade into the collagen layer. In vivo, type V collagen, expressed in the lower stromal layer and the surface of the myometrium, may play a role to maintain the early embryo in the decidual compartment. EGTA inhibited the attachment of the blastocysts. Anti-laminin antibody and RGDS peptide, attachment domain of laminin, also inhibited the implantation. These findings suggested that the Ca2+ dependent process was necessary for the attachment between the trophoblasts and the endometrial cells, and then the implantation process was triggered after the attachment to the laminin in basement membrane. The endometrial tissue, obtained from the infertile patient, was cultured on the basement membrane extract with serially obtained maternal serum. Addition of the maternal serum after proper administration of pFSH, high estrogen conditions were made in the culture dishes. These conditions increased the height of the glandular structure and relatively decreased the area of the surface epithelium. Decreased the area of surface epithelium affected the rate of the attachment. Endometrial cell culture system associated with functional and morphological characteristics was established. Serial observation of the endometrial cells in this system revealed the rabbit and the human implantation process, and the embryo-endometrial interaction.(ABSTRACT TRUNCATED AT 400 WORDS)

摘要

着床被认为是母体与胚胎之间相互作用且同步的过程,然而,早期着床过程的机制仍未阐明。因此,体外受精和胚胎移植的有效性仍然较低。结果;免疫组织化学检测显示,IV型胶原蛋白和其他细胞外基质成分主要在上皮层下方被检测到。I型和III型胶原蛋白在基质层中呈弥漫性分布。在基质下层和浅表肌层中,检测到了V型胶原蛋白。人子宫内膜上皮细胞在腺体形成后进行了再上皮化。兔子宫内膜细胞在褶皱形成后也进行了再上皮化。基质细胞侵入到褶皱的内层。添加到培养基中的雌激素刺激了腺体的形成。在雌激素预处理后给予孕酮并不影响腺体的形成,然而,浅表上皮和再上皮化区域的增殖增加。兔囊胚成功附着并植入到重建的子宫内膜中。植入胚胎的发育在形态上是正常的。人培养的滋养层细胞附着、侵入并穿透细胞外基质成分。另一方面,使用包被有V型胶原蛋白的培养皿,滋养层细胞能够侵入基质层,然而,V型胶原蛋白层不允许滋养层细胞侵入到胶原蛋白层。在体内,在基质下层和肌层表面表达的V型胶原蛋白可能在将早期胚胎维持在蜕膜腔中发挥作用。乙二醇双乙醚二胺四乙酸(EGTA)抑制了囊胚的附着。抗层粘连蛋白抗体和层粘连蛋白的附着结构域RGDS肽也抑制了着床。这些发现表明,Ca2+依赖过程对于滋养层细胞与子宫内膜细胞之间的附着是必要的,然后在附着于基底膜中的层粘连蛋白后触发着床过程。从不孕患者获取的子宫内膜组织在含有连续获取的母体血清的基底膜提取物上进行培养。在适当给予促卵泡生成素(pFSH)后添加母体血清,在培养皿中创造了高雌激素条件。这些条件增加了腺体结构的高度并相对减小了表面上皮的面积。表面上皮面积的减小影响了附着率。建立了与功能和形态特征相关的子宫内膜细胞培养系统。对该系统中子宫内膜细胞的连续观察揭示了兔和人的着床过程以及胚胎 - 子宫内膜相互作用。(摘要截取自400字)

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