To start, incubate fertilized chicken eggs in a humidified incubator at 38 degrees Celsius. Then, place a stage 28 to 32 chicken embryo in a 60-millimeter Petri dish filled with HBSS. After dissecting out the dorsal embryo skin and head, gently pull the limb buds to reposition the ventral side of the body downwards.
Now, along the two sides of the embryo body, make an anterior to posterior incision. Grip the limb buds longitudinally with a pair of forceps to stabilize the body. Use a watchmaker's forceps to carefully peel the skin from the neck down to the tail region.
Use sharp scissors to delicately remove any remaining subcutaneous tissues still attached to the peeled skin and smooth the skin edges. Next, add two milliliters of supplemented DMEM to each well of a six-well dish. After adding antibiotics to the wells, place a sterile tissue culture insert inside the well, ensuring the medium surrounds the exterior of the culture insert.
Then move the skin onto a spatula while immersed in HBSS to prevent creasing and folding. Once done, transfer the excised skin to a dish containing HBSS. Slowly slide the skin off the spatula onto the culture insert without folding.
Using a 200-microliter pipette, remove any excess HBSS from the insert. Finally, incubate the skin explant cultures at 37 degrees Celsius with a blend of 5%carbon dioxide and 95%air. Replace the medium every two days.
The feather primordia developed into short feather buds after two days in culture. and long feather buds developed after four days in culture. The dermal placodes were more mature in the midline than at the sides.
