| Budget Amount *help |
¥2,100,000 (Direct Cost: ¥2,100,000)
Fiscal Year 1996: ¥1,100,000 (Direct Cost: ¥1,100,000)
Fiscal Year 1995: ¥1,000,000 (Direct Cost: ¥1,000,000)
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| Research Abstract |
Endometrial cells play crucial roles in the implantation of blastocysts, which proceeds primarily under the influence of sex steroid hormones. The actions of these hormones have been investigated using monolayr cultures consisting of endometrial cells and embryonic cells. The present study developed a novel three-dimensional endometrial cell culture system producing an in vivo-like microenvironment. A three-dimensional tissue-like mass of human endometrial stromal cells (spheroid) was prepared utilizing a conjugate of collagen and a thermo-responsive polymer, poly-N-isopropyl acrylamide (PNIP AAm), as a substrate for cell adhesion. The endometrial stromal cells in the spheroid responded well to progesterone and reorganized their morphological arrangement to form "nests". They expressed prolactin, desmin, and connexin43, indicating that progesterone induced differentiation in endometrial stromal cells. The endometrial spheroid is a simple and experimentally controllable culture that mim
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icsin vivo responses, and, therefore, is useful for the analysis of cell-to-cell interactions during the implatation process. At least four parameters are critical for determining the extent of differentiation in cultured cells 1) cell shape, 2) cell density for cell-cell communication, 3) cell-ECM interaction, 4) interaction between epithelial and mesenchymal cells related to the paracrine control. However, no culture system can produce optimal conditions simultaneously. Moreover, in order to reconstitute the sex steroid hormone-responsive tissue equivalent, stromal cells must not only underlie the epithelium in the cultures, but also be functional in responding to the sex steroid hormones. Using new artificial substrata, a novel culture method in which not only stromal but also epithelial cells can respond to sex steroid hormones could be developed in a near future. Progress in cell technology and biomaterials for the scaffold of cells also could enhance the development of the three dimensional culture technology and could produce a culture system in which cells can respond well to sex steroid hormones. Less
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