Establishing Villous and Decidual Organ Cultures as Ex Vivo Models of Human Maternal-Fetal Interface
Trascrizione
Begin with placental villous and decidual parietalis specimens.
Dissect the villous — a well-vascularized tissue containing extravillous trophoblast, or EVT, cell columns.
Transfer the villous over an extracellular matrix, ECM-coated transwell placed in a tissue culture well.
Take the next specimen; microdissect the decidua — a uterine tissue surrounding the developing fetus.
Place the decidua over another ECM-coated transwell.
Add a suitable medium to the tissue culture wells to maintain humidity, while keeping the transwells devoid of medium.
Without a medium, the EVT cell columns secrete proteinases that cleave ECM proteins.
This initiates ECM remodeling, creating space for the EVT cells to invade and firmly adhere to the matrix.
Meanwhile, the other cells of the villous and decidua attach to the ECM by activating integrin receptors, which cluster to bind cooperatively to the matrix.
These processes strengthen the cell-to-ECM adhesion, securely embedding the tissues.
Finally, add tissue-specific media to the transwells for culture growth.
