In vitro 着床モデルを用いた受精卵着床機構の研究(<シンポジウム>妊娠の成立機構 : 胚発生から着床まで)

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Implantation is thought to be interactive and synchronized process between mother and embryo, however, the mechanisms of the early implantation process remain unelucidated. Therefor, 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 Ca^<2+> 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 functinal 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. It is suggested that the implantation process was controlled by the communication between trophoblast and endometrial cell, and this process was divided into 4 stages, Ca^<2+>-dependent cell attachment, trophoblast fusion into superficial epithelium, penetration into basement membrane, and invasion into stromal layer and embedding. Synchronized culture of the endometrial cell obtained from infertile patient in the treatment cycle provided the modalities to evaluate the uterine capacity of patient for implantation in vitro. These findings help us to decide the timing of implantation, the protocol of the hormonal support and clinical significance of the oocyte cryopreservation. Furthermore, these studies suggested that the development of the in vitro implantation system will make advance in both clinical and fundamental fields of the reproductive physiology.
社団法人日本産科婦人科学会の論文
1992-08-01

In vitro 着床モデルを用いた受精卵着床機構の研究(<シンポジウム>妊娠の成立機構 : 胚発生から着床まで)

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