Establishing Villous and Decidual Organ Cultures as Ex Vivo Models of Human Maternal-Fetal Interface

Using sterile technique in a tissue culture hood, thaw extracellular matrix files on ice, then, dilute the warmed extracellular matrix at a 1 to 1 ratio with ice cold collection medium. Mix the suspension with pipetting, taking care not to induce bubbles.

Next, place transwell inserts with 0.4-micron pores into individual wells of a 6-well tissue culture plate and coat each insert with 100 microliters of the extracellular matrix suspension. Place the plate on ice, and rinse the specimens 2 times in collection medium, transferring the tissue to a sterile petri dish under a dissecting microscope after the second wash.

The specimens contain 2 distinct types of decidual tissues; decidua parietalis and decidua capsularis. Using sterile springer dissecting scissors and forceps, microdissect the decidua parietalis into 3-cubic-millimeter pieces, using the forceps to tease away any clotted maternal blood and use forceps to transfer 3 or 4 pieces of decidua into each transwell.

Identify the well-vascularized villi with prominent extravillous trophoblast cell columns and use forceps to transfer 3 or 4 villous trees into each transwell. Adjust the branches, so that the trees are positioned flat atop the extracellular matrix and separate any clumped branches. Then, add 1 milliliter of collection medium to the bottom of each well and incubate the cultures overnight at 37 degrees Celsius in 5% carbon dioxide. 12 to 16 hours later, add 1 milliliter of the appropriate culture medium to the top of each transwell.