Gut Churning

The discovery of an intestinal stem cell marker fuels an ongoing debate over the cells' location and properties.

GFP-labeled Lgr5-positive cells in the crypt base of the mouse intestine
Courtesy of Nick Barker and Hugo Snippert

Mammalian intestinal epithelium is one of the most swiftly self-renewing tissues in the body, turning over completely every 3 to 5 days. Because of the absence of reliable stem cell markers, however, researchers have argued for decades about the identity and location of the stem cells that fuel this growth capacity.

In the intestinal epithelium, cells proliferate in glandular pockets termed the crypts of Lieberkühn. In the 1970s, two competing theories emerged for where in the crypt these all-important, self-renewing cells abide. The predominant idea, put forth by Chris Potten at the Paterson Institute for Cancer Research in Manchester, United Kingdom, and a cofounder of Epistem, an epithelial stem cell company,...

In 2007, Hans Clevers's group at the Hubrecht Institute for Developmental Biology and Stem Cell Research in Utrecht used a combination of inducible gene knock-ins and lineage tracing to identify a gene that serves as an intestinal stem cell marker.3 Cells positive for the gene, leucine-rich, repeat-containing G proteincoupled receptor 5 (Lgr5), localized to the region where CBC cells reside, and generated all the lineages seen in the intestinal and colon epithelium. These Lgr5-positive cells divide once per day—a point of controversy, since many researchers hold that stem cells divide only rarely (on the order of weeks or months), in order to maintain the stability of their genome.

The Nature study, this month's Hot Paper, provided the first genetic marker for intestinal cells with distinct stem cell behavior, and revived Leblond's hypothesis regarding where to find them. "For us, this is old news," says Matthew Bjerknes of the University of Toronto, who, along with Leblond's student Hazel Cheng has continued working on the CBC hypothesis. "But it has opened the debate widely again."

Location, location, location

Clevers quickly followed his 2007 Lgr5 paper with several others supporting that Lgr5-positive CBC cells were, in fact, stem cells. In one, a March Nature paper, his group purified and cultured Lgr5- positive cells in the presence of a Wnt factor, and the cultures formed "organoids" with distinct crypt-villus structures.4 Culturing gut epithelium has been notoriously challenging, and growing whole segments of gut in culture "is spectacular," says Riccardo Fodde of the Erasmus Medical Center in Rotterdam. "It is unbelievable almost."

Meanwhile, the debate over intestinal stem cells' identity and location is far from over. Last summer, Mario Capecchi at the University of Utah School of Medicine reported that Bmi1, already implicated in self-renewal in neurons and hematopoietic cells, marks intestinal stem cells, and pinpointed them to approximately the +4 position.5 More indirect evidence from Linheng Li's lab at the Stowers Institute for Medical Research in Kansas City, Missouri, implicates yet another gene in intestinal stemness that localizes around +4.6

Courtesy of Hans Clevers

According to Potten, those two lines of evidence definitively support his model. Clevers's 2007 study showed a few Lgr5-positive cells in position +4, and Potten says those cells—which would also have been present in the purified cells of the "organoid" study—are the intestinal stem cells. (Lgr5's function is unknown; Potten speculates the labeled cells at the base of the crypt are precursors to another nonstem-cell type.)

Clevers, however, notes Li's marker was never functionally verified, and insists that for stem cells, Lgr5 trumps Bmi1. "On its own," he says, Lgr5 "makes a black and white distinction between a stem cell and a daughter cell. And I don't know any other marker that does that." A recent gene expression study from his lab showed his Lgr5-positive cells also expressed Bmi1. The two labeled populations also have the same kinetics, he adds: just like Lgr5-positive cells, Bmi1-positive cells fill the crypt with lots of cells in 3 or 4 days.7 Indeed, says Clevers, he is convinced his group and Capecchi's "are actually tracing the same cells."

Beyond the gut

Intestine isn't the only tissue to hold putative stem cells carrying Lgr5: Elaine Fuchs at The Rockefeller University notes that the gene has consistently come up in screens of hair follicle stem cells. The biology of hair follicle stem cells is better understood than intestine and other structures, so researchers can use that knowledge to better understand Lgr5's role. "I think what's exciting is that we can now draw on each other's science, and that always propels a field forward," she says.

Clevers agrees that in comparing markers in different systems, "we hope to see a minimum program of stemness." His group is working on teasing out the function of Lgr5 to better understand what precisely it's marking. In addition, they have found Lgr5- positive cells in the stomach, brain, retina, mammary gland, kidney, and pancreas. Except for the mammary gland, he adds, "in all cases, they sit in different places than where people had postulated them to be"—and by extension, mark different populations than past studies have fingered as stem cells.

With the advent of reliable stem cell markers, the debate regarding location may now be irrelevant. "We can ask ourselves, does it really matter anymore?" says Bjerknes. "Now we have tools in addition to simple morphology. We have these markers, let's just push forward and see what these cells can do."

Data derived from the Science Watch/Hot Papers database and the Web of Science (Thomson ISI) show that Hot Papers are cited 50 to 100 times more often than the average paper of the same type and age.

Correction (July 7): A previous version of this article incorrectly stated Charles Philippe Leblond's university affiliation as the University of Toronto. In fact, he was based at McGill University in Montreal. The Scientist regrets the error.

N. Barker et al., "Identification of stem cells in small intestine and colon by marker gene Lgr5," Nature, 449:1003–8, 2007. (Cited in 150 papers)
1. C.D. Potten et al., "Continuous labeling studies on mouse skin and intestine," Cell Tissue Kinet, 7:271–83, 1974.
2. H. Cheng and C.P. Leblond, "Origin, differentiation and renewal of the four main epithelial cell types in the mouse small intestine. I. Columnar cell," Am J Anat, 141:461–79, 1974.
3. N. Barker et al., "Identification of stem cells in small intestine and colon by marker gene Lgr5," Nature, 449:1003–7. 2007.
4. T. Sato et al., "Single Lgr5 stem cells build crypt-villus structures in vitro without a mesenchymal niche," Nature, March 2009 [EPUB].
5. E. Sangiorgi and M. Capecchi, "Bmi1 is expressed in vivo in intestinal stem cells," Nat Genet, 40:915–20, 2008.
6. X.E. He et al., "PTEN-deficient intestinal stem cells initiate intestinal polyposis," Nat Genet, 39:189–98, 2007.
7. LG van der Flier et al., "Transcription factor achaete scute-like 2 controls intestinal stem cell fate," Cell, 136:903–12, 2009.

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