A novel method of isolating high quality RNA from Kupffer cells

Apr 17, 2009

Kupffer cells, resident tissue macrophages that line the liver sinusoids, play a key role in modulating inflammation in a number of experimental models of liver injury. Since Kupffer cells represent only a small portion of the entire liver cell population, greatly outnumbered by the parenchymal cells, Kupffer cell isolation faces major technical obstacles. Laser capture microdissection (LCM) offers a method of isolating a single cell type from specific regions of tissue sections.

A research team led by Dr Stephen H Gregory from United States outlines isolation of Kupffer using LCM. This will be published on April 14, 2009 in the .

LCM is an essential approach used in conjunction with molecular analysis to study the functional interaction of cells in their native tissue environment. The process of labeling and acquiring cells by LCM prior to mRNA isolation can be elaborate, thereby subjecting the RNA to considerable degradation. Kupffer cell labeling was achieved by injecting India ink intravenously, thus circumventing the need for in vitro staining. The significance of this novel approach was validated using a cholestatic liver injury model.

They found that mRNA extracted from the microdissected displayed marked increases in colony stimulating factor-1 receptor and Kupffer cell receptor message expression, which demonstrated Kupffer cell enrichment. by Kupffer cells derived from bile-duct-ligated, versus sham-operated, mice was compared. Microarray analysis revealed a significant (2.5-fold, q value < 10) change in 493 genes. based on this fold-change and a standardized pubmed search, 10 genes were identified that were relevant to the ability of kupffer cells to suppress

The methodology outlined herein provides an approach to isolating high quality RNA from Kupffer cells, without altering the tissue integrity.

More information: Gehring S, Sabo E, San Martin ME, Dickson EM, Cheng CW, Gregory SH. Laser capture microdissection and genetic analysis of carbon-labeled Kupffer . World J Gastroenterol 2009; 15(14): 1708-1718 http://www.wjgnet.com/1007-9327/15/1708.asp

Source: World Journal of Gastroenterology (news : web)

Explore further: Researcher develops, proves effectiveness of new drug for spinal muscular atrophy

add to favorites email to friend print save as pdf

Related Stories

How did glycine significantly decrease liver injury?

Oct 31, 2008

The nonessential amino acid glycine has been shown to be anti-inflammatory in several animal injury models. Recent studies demonstrated that dietary glycine protected both the lung and liver against lethal doses of endotoxin ...

What is the role of the omentum in regenerating the liver?

Mar 09, 2009

In their recent work to be published on March 7, 2009 in the World Journal of Gastroenterology, Dr. Singh and his colleagues from the Cook County Hospital in Chicago (USA) first activated the omentum using a foreign body t ...

Recommended for you

Cellular protein may be key to longevity

17 hours ago

Researchers have found that levels of a regulatory protein called ATF4, and the corresponding levels of the molecules whose expression it controls, are elevated in the livers of mice exposed to multiple interventions ...

Gut bacteria tire out T cells

20 hours ago

Leaky intestines may cripple bacteria-fighting immune cells in patients with a rare hereditary disease, according to a study by researchers in Lausanne, Switzerland. The study, published in The Journal of Experimental Me ...

T-bet tackles hepatitis

20 hours ago

A single protein may tip the balance between ridding the body of a dangerous virus and enduring life-long chronic infection, according to a report appearing in The Journal of Experimental Medicine.

User comments : 0