Enrichment for murine keratinocyte stem cells based on cell surface phenotype

Tani, Hiroshi; Morris, Rebecca J.; Kaur, Pritinder

doi:10.1073/pnas.97.20.10960

Cited by 339 publications

(317 citation statements)

References 31 publications

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“…2I-L). It has been noted that stem cells isolated from other tissues such as the skin and hair follicle [43][44][45] have smaller cell sizes than their more differentiated progeny. The SP cell frequency in rat corneal epithelium (4.6%) was also much higher than in the limbus, as well as results reported for several tissues and organs [13,16,20,23], and the presence of corneal epithelial SP cells in rat eyes were contrary to our previous results for both human [23] and rabbit [24].…”

Section: Discussionmentioning

confidence: 99%

Rat limbal epithelial side population cells exhibit a distinct expression of stem cell markers that are lacking in side population cells from the central cornea

et al. 2005

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The side population (SP) phenotype is shared by stem cells in various tissues and species. Here we demonstrate SP cells with Hoechst dye efflux were surprisingly collected from the epithelia of both the rat limbus and central cornea, unlike in human and rabbit eyes. Our results show that rat limbal SP cells have a significantly higher expression of the stem cell markers ABCG2, nestin, and notch 1, compared to central corneal SP cells. Immunohistochemistry also revealed that ABCG2 and the epithelial stem/progenitor cell marker p63 were expressed only in basal limbal epithelial cells. These results demonstrate that ABCG2 expression is closely linked to the stem cell phenotype of SP cells.

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Section: Discussionmentioning

confidence: 99%

Rat limbal epithelial side population cells exhibit a distinct expression of stem cell markers that are lacking in side population cells from the central cornea

et al. 2005

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“…Cell surface receptor molecules are particularly useful for isolating and maintaining specific cell subsets in vitro for investigational studies (Kaur and Li, 2000). However, reliance on cell adhesion molecules alone is inadequate for the purification of stem cell candidates, because almost all basal keratinocytes express, to varying degrees, these molecules which are essential for many different biological processes (Peltonen et al, 1989;Carter et al, 1990;Dowling et al, 1996;Hodivala-Dilke et al, 1998;Tani et al, 2000). Kaur and colleagues have identified and enriched putative stem cells from epidermal keratinocytes (Peltonen et al, 1989;Li et al, 1998).…”

Section: Introductionmentioning

confidence: 99%

Neurotrophin receptor p75NTR characterizes human esophageal keratinocyte stem cells in vitro

et al. 2003

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We report here that human esophageal keratinocyte stem cells are characterized by the expression of the lowaffinity neurotrophin receptor p75 NTR and differentially expressed cell adhesion molecules, the b1 and b4 integrins. The candidate stem cells could be fractionated from keratinocytes as a minor cell subset by means of immunocytochemical cell sorting based on the different levels of expression of these cell surface molecules. Flow cytometric analysis revealed that this minor cell subset retained a relatively slow-cycling phenotype in vitro. These cells expressed low levels of involucrin and cytokeratin 13, indicating that the p75 NTR -positive cell subset is immature relative to the other predominant subpopulations coexpressing b1 integrin at higher levels. The p75 NTR -positive cell subset was crucial for achieving longevity and the greatest output of keratinocytes comprising all distinguishable subpopulations in vitro. This process was associated with self-renewal and selfamplification of the p75 NTR -positive cell subset. These findings strongly implicate p75 NTR as a stem cell marker, which will be valuable for prospectively investigating stem cell regulation in association with different biological processes including neoplastic transformation of regenerative epithelia.

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“…2 FACS analyses revealed that B18% of total epidermal cells were integrin b1 + (b1 + ) and 6.5% were CD71 + compared with the isotype control. The 14.1% of primary keratinocytes that were b1 + CD71 À cells were efficiently enriched by FACS to 91.2% purity (Figure 1b).…”

Section: Resultsmentioning

confidence: 99%

“…Candidate murine keratinocyte stem cells express high levels of b1 integrin and low levels of the transferrin receptor, CD71 (b1 integrin + CD71 À ). 2 Additional markers of hair follicle stem cells include the combination of high expression of integrin a6 (a6+), low levels of CD71 (CD71 À ) and high CD34 levels. 2 Hair growth, differentiation and regeneration involve several signaling pathways, 3 including Wnt signaling, an important pathway in hair growth.…”

Section: Introductionmentioning

confidence: 99%

“…2 Additional markers of hair follicle stem cells include the combination of high expression of integrin a6 (a6+), low levels of CD71 (CD71 À ) and high CD34 levels. 2 Hair growth, differentiation and regeneration involve several signaling pathways, 3 including Wnt signaling, an important pathway in hair growth. 4 Telomerase reverse transcriptase (TERT), which is expressed during anagen, upregulates Wnt-signaling pathway.…”

Section: Introductionmentioning

confidence: 99%

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The use of polyethylenimine–DNA to topically deliver hTERT to promote hair growth

et al. 2011

Gene Ther

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The present study investigates the efficacy of polyethylenimine (PEI)-DNA complex that expressed human telomerase reverse transcriptase (hTERT) to transfect hair follicle stem cells and produce sufficient hTERT to stimulate hair growth. Transfection with pLC-hTERT-DNA-PEI complex (D+P group) in vitro induced expression of proliferating cell nuclear antigen in 35.8% of the purified stem cell population, suggesting enhanced cell proliferation. In vivo transfection efficiency of rat dorsal skin was determined by staining for b-gal activity. Cells positive for b-gal were located in the bulge region and dermal sheath of hair follicles. The follicles in the hTERT-transfected region entered anagenon day 15 after transfection, whereas non-transfected (Neg) controls remained in telogen. The similar effect was observed in 50-day-old rat dorsal skin. D+P group displayed a specific expression of hTERT and sufficient to initiate a transition to the anagen phase and promote new hair synthesis 18 days after the transfection. hTERT promoted follicle neogenesis following wounding. In all, 60 days after wounding, tissues of the D+P group showed more newly regenerating hair follicles (83±52 regenerated follicles per rat) in contrast to control group tissues (15±15 regenerated follicles per rat). These studies provide a potential approach for gene therapy of skin disease.

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Enrichment for murine keratinocyte stem cells based on cell surface phenotype

Cited by 339 publications

References 31 publications

Rat limbal epithelial side population cells exhibit a distinct expression of stem cell markers that are lacking in side population cells from the central cornea

Rat limbal epithelial side population cells exhibit a distinct expression of stem cell markers that are lacking in side population cells from the central cornea

Neurotrophin receptor p75NTR characterizes human esophageal keratinocyte stem cells in vitro

The use of polyethylenimine–DNA to topically deliver hTERT to promote hair growth

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