Human primary mesothelial cells, or HPMCs, line many visceral organs like the omentum or the layer of fatty tissue present in the peritoneal cavity.
To isolate the mesothelial cells, begin by immersing a human omentum specimen in a suitable buffer to remove the excess red blood cells or RBCs. Next, place the omentum in a glass dish and mince it into pieces. This dissociates the tissue, releasing HPMCs along with the contaminating RBCs.
Transfer the pieces into a conical tube and allow them to remain still for the desired time. The minced tissue rich in low-density fat rises to the top, while the dense cellular components settle at the bottom. Next, aspirate the liquid at the bottom, containing HPMCs and RBCs, into a fresh tube.
Centrifuge to collect the cells. Remove the buffer-containing supernatant. Resuspend the pellet in a mesothelial growth medium and incubate for the desired duration. The media facilitates the selective growth of HPMCs over RBCs.
Observe the cells using a light microscope to confirm the growth of cuboidal-shaped human mesothelial cells.