Regeneration of cell culture substrata using bovine uterine and placentomal extra-cellular matrix and remodeling of endometrial functions

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The aim of this study was to explore the way to use of a new extra-cellular matrix (ECM) substrata from bovine utero-placental tissues and the regulatory mechanisms for endometrial functions regarding the interaction between trophoblast cells and endometrium around implantation for establishing a mimic in vitro research tool of bovine implantation. Urea buffer solution and sodium deoxycholate were effective to extract ECM from utero-placental tissues, and these extracts stimulated the growth of bovine endometrial fibroblasts. Fibroblasts well grew and developed a firm and adaptable cell-sheet on ECM extracted by sodium deoxycholate. The sheet may be an utilizable substratum for generating endometrial-like multi-cellular structure. Microarray and in situ hybridization analysis suggested that some molecules, like placental lactogen, prolactin-related proteins -1 (PRP1), PRP5, pregnancy-associated glycoprotein 7, cathepsin K, insulin growth factor binding protein, took a crucial role for implantation, and ECM degradation related enzymes; membrane type I-MMP and extracellular matrix metalloproteinase inducer. EMMPRIN, were participated in the remodeling of endometrium and implantation in bovine. PRP-1 specifically bound to basement membrane component, type IV collagen. The molecule may be a key factor for adhesion and implantation between early embryo and endometrial epithelium in bovine. Co-culture of trophoblast cell line (BT-1 cell) and endometrial fibroblasts suggested that trohoblast cell produced some specific molecule to stimulate fibroblast cell growth efficiently. Overall the substrata from bovine utero-placental ECM may retain an useful feature for regeneration of multi-cellular structure using cells and matrix components in vitro, and also the information of ECM degradation enzymes and adhesion molecules in this study may accelerate establishing in vitro endometrial models, however, more sophisticated arrangements need for establishing them.