How do you isolate mouse peritoneal macrophages?
How do you isolate mouse peritoneal macrophages?
Using a scissors and forceps cut the outer skin of the peritoneum and gently pull it back to expose the inner skin lining the peritoneal cavity. Inject 5 ml of ice cold PBS (with 3% FCS) into the peritoneal cavity using a 27g needle. Push the needle slowly in the peritoneum being careful not to puncture any organs.
What part of the mice can we isolate macrophages from for the phagocytosis experiment?
mouse peritoneal cavity
Generally, macrophages isolated from the mouse peritoneal cavity will be mature quiescent macrophages (Cohn, 1978; Fortier et al., 1982).
What was the purpose of using cells from the peritoneal cavity?
The peritoneal cavity in mammals contains different immune cell populations crucial for innate immune responses. An efficient isolation method is required for biochemical and functional analyses of these cells.
Where is the peritoneal cavity in mice?
The peritoneal cavity is a membrane-bound and fluid-filled abdominal cavity of mammals, which contains the liver, spleen, most of the gastro-intestinal tract and other viscera. It harbors a number of immune cells including macrophages, B cells and T cells.
What is the function of peritoneal membrane?
The peritoneal membrane is the smooth, transparent membrane that lines the abdominal cavity and contains the internal organs of the abdomen and pelvis, such as the stomach and large intestine. The peritoneal membrane helps to protect and separate the internal structures of the abdomen and pelvis.
Where does peritoneal fluid come from?
Peritoneal fluid is the fluid from the peritoneal cavity, a space between the wall of the abdomen and the organs inside.
How long can you keep macrophages in culture?
In my experience they usually last about 3 days after the 7 days of differentiation. I was able to follow macrophages for 12 days after PBMC plating if no growth factor was added and also currently have them for 1 months with GM-CSF stimulation (30 ng/ml). M-CSF may keep them alive for a long time as well.
