This article details a method for isolating peritoneal fat depots in female mice, which may provide a microenvironment for cancer cell colonization during metastasis. The procedure involves surgical dissection to extract various fat tissues, including omental, gonadal, uterine, mesenteric, and splenoportal fat.
The adipose tissue or the fat depots primarily present in the omental, mesenteric, uterine, gonadal, and splenoportal regions of the peritoneum may provide a suitable microenvironment for cancer cell colonization during metastasis.
To isolate these peritoneal fat depots, begin by taking a euthanized female mouse. Make a midline surgical incision through the peritoneal wall to expose its internal organs. Now, locate the omental fat present over the stomach and pancreatic spleen complex and extract it. Excise the gonadal fat surrounding the ovaries. Further, remove the uterine fat enveloping the uterine horns. Subsequently, cut the junction between the small intestine and pylorus to release the mesenteric fat attached to the intestine. Finally, extract the thin band of splenoportal fat connecting the distal end of the spleen with the pancreas.
Maintain the dissected fat tissues in a suitable chilled buffer to preserve tissue physiology. Analyze these peritoneal fat tissues to assess the presence of colonized metastatic cancer cells.
As harvesting peritoneal fat depots constitutes internal organ removal, make a midline incision close to the inguinal papillae through the peritoneal wall to expose the internal organs. To excise the omental fat, expose the omental pancreatic complex by extending the spleen from the peritoneal cavity with forceps.
For the gonadal fat, use forceps to lift the gonadal fat surrounding the ovaries and excise it by cutting tissue connections. Immediately place the tissues in ice-cold PBS. Next, remove the uterine fat by using forceps to lift the uterine fat surrounding the uterine horns and excise by cutting the tissue connections before immediately placing the tissues in ice-cold PBS.
When obtaining the mesenteric fat, cut the junction between the small intestine and the pylorus. Use forceps to firmly grip the free end of the small intestine and slowly peel it away from the mesenteric fat. Release the mesenteric fat from the mesenteric root using dissecting scissors. Then, immediately place the tissues in ice-cold PBS.
Finally, to remove the splenoportal fat, lift the distal end of the spleen using forceps to expose the thin fatty band of tissue connecting the hilum of the spleen to the pancreas. Excise the splenoportal fat by first releasing it from the pancreas and then carefully dissecting it from the spleen. Then, immediately place the tissues in PBS.
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