Proximal Culture System: An In Vitro Culture Technique to Study Paracrine Signaling Between Cells
Transcrição
In paracrine signaling, cells produce and secrete signaling molecules that induce responses in neighboring cells. To mimic paracrine signaling, begin by preparing single-cell suspensions of ovarian cancer cells and mesothelial cells in their appropriate medium.
Next, take an inverted porous membranous insert in a sterile culture dish. Pipet the cancer cell suspension onto the bottom surface of the insert membrane to form a liquid dome. Incubate to allow the cells to settle down and attach to the membrane.
Invert the insert to drain the media. Transfer the insert into a fresh culture dish containing growth media to nourish the cancer cells adhered to the bottom surface of the membrane. Now, add the mesothelial cell suspension into the insert well. Incubate to allow the cells to adhere to the opposite side of the membrane, denying any direct contact between the two cell types.
During culture, both cell types secrete signaling factors. These molecules diffuse through the porous membrane towards the neighboring cells. They bind to specific cell receptors and induce complex signaling cascades in both cell types.
