Cecal ligation and puncture

Abstract

The cecum contains a high concentration of microbes, which are a combination of Gram-negative and Gram-positive flora. These bacteria range from anaerobic to facultative aerobic to aerobic organisms. In the procedure described in this unit, the ligation of the cecum produces a source of ischemic tissue as well as polymicrobial infection. This combination of ischemic/necrotic tissue and microbial infection distinguishes this multifactorial model from a number of other bacterial sepsis models, including but not limited to: bacteremia secondary to intravenous or intraperitoneal administration; fecal administration or intraperitoneal administration of fecal or bacterial plugs; colonic stents; and bacterial abscess formation.

Figure 1

Midline incision through the skin.

Figure 2

Exteriorization of the cecum.

Figure 3

Ligation and needle puncture of the cecum. A. One centimeter of cecum has been suture ligated. B. The ligated cecum has been punctured through and through with hollow bore needle.

Figure 4

Closure of the abdomen. A. Closure of abdominal musculature. B. Closure of skin with auto-stapler.

Figure 5

Injection of saline/buprenorphine into the scruff of the mouse.

Figure 6

Peritonitis/bacteremia and the immune response after CLP. A. Blood and Peritoneal Washes cultured on Blood Agar [14] B. Cytokine expression during sepsis [14] C. Peripheral white blood cells (WBCs) in Sham and CLP animals [14].

Figure 7

CLP survival curve using different gauge needles for puncture (Unpublished data).