Human limbal epithelium contains side population cells expressing the ATP‐binding cassette transporter ABCG2

Watanabe, Katsuhiko; Nishida, Kohji; Yamato, Masayuki; Umemoto, Terumasa; Sumide, Taizo; Yamamoto, Kazuaki; Maeda, Naoyuki; Watanabe, Hitoshi; Okano, Teruo; Tano, Yasuo

doi:10.1016/j.febslet.2004.03.064

Cited by 196 publications

(81 citation statements)

References 40 publications

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“…Several studies have provided support for this proposal (Scharenberg et al, 2002;Lechner et al, 2002;Kim and Morshead, 2003;Alison, 2003;Lassalle et al, 2003;Bhattacharya et al, 2003;Summer et al, 2003). Preliminary reports on the presence of SP cells in the limbal zone have been published (Watanabe et al, 2004;Wolosin et al, 2004) We describe now the existence of ABCG2-dependent SP cells in the human and rabbit ocular surface epithelia and demonstrate that these cells display many features that are consistent with a stem cell phenotype.…”

Section: Introductionmentioning

confidence: 54%

Ocular surface epithelia contain ABCG2-dependent side population cells exhibiting features associated with stem cells

Budak

Alpdoğan

Zhou

et al. 2005

Journal of Cell Science

209

145

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When cell populations are incubated with the DNA-binding dye Hoechst 33342 and subjected to flow cytometry analysis for Hoechst 33342 emissions, active efflux of the dye by the ABCG2/BCRP1 transporter causes certain cells to appear as a segregated cohort, known as a side population (SP). Stem cells from several tissues have been shown to possess the SP phenotype. As the lack of specific surface markers has hindered the isolation and subsequent biochemical characterization of epithelial stem cells this study sought to determine the existence of SP cells and expression of ABCG2 in the epithelia of the ocular surface and evaluate whether such SP cells had features associated with epithelial stem cells. Human and rabbit limbal-corneal and conjunctival epithelial cells were incubated with Hoechst 33342, and analyzed and sorted by flow cytometry. Sorted cells were subjected to several tests to determine whether the isolated SP cells displayed features consistent with the stem cell phenotype. Side populations amounting to <1% of total cells, which were sensitive to the ABCG2-inhibitor fumitremorgin C, were found in the conjunctival and limbal epithelia, but were absent from the stem cell-free corneal epithelium. Immunohistochemistry was used to establish the spatial expression pattern of ABCG2. The antigen was detected in clusters of conjunctival and limbal epithelia basal cells but was not present in the corneal epithelium. SP cells were characterized by extremely low light side scattering and contained a high percentage of cells that: showed slow cycling prior to tissue collection; exhibited an initial delay in proliferation after culturing; and displayed clonogenic capacity and resistance to phorbol-induced differentiation; all features that are consistent with a stem cell phenotype.

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Section: Introductionmentioning

confidence: 54%

Ocular surface epithelia contain ABCG2-dependent side population cells exhibiting features associated with stem cells

Budak

Alpdoğan

Zhou

et al. 2005

Journal of Cell Science

209

145

View full text Add to dashboard Cite

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“…Conceptually, they may form a component of the molecular mechanisms by which long-lived stem cells reduce the potential for genomic damage over their extended lives, and their expression has been correlated with stem cell activity [90] . ABCG2 expression in the limbus is one such example and cells expressing this marker can be isolated as a "side population" by fluorescenceactivated cell sorting (FACS) [3,57,[91][92][93][94][95] . However, as noted above, some of the ABCG2-positive, labelretaining cells with a high nucleus to cytoplasm ratio cells in the rat limbus have been identified as Langerhans cells rather than epithelial stem cells [56] .…”

Section: Stem Cell Markers and Phenotypementioning

confidence: 99%

Evaluating alternative stem cell hypotheses for adult corneal epithelial maintenance

West¹,

Dora²,

Collinson

2015

WJSC

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In this review we evaluate evidence for three different hypotheses that explain how the corneal epithelium is maintained. The limbal epithelial stem cell (LESC) hypothesis is most widely accepted. This proposes that stem cells in the basal layer of the limbal epithelium, at the periphery of the cornea, maintain themselves and also produce transient (or transit) amplifying cells (TACs). TACs then move centripetally to the centre of the cornea in the basal layer of the corneal epithelium and also replenish cells in the overlying suprabasal layers. The LESCs maintain the corneal epithelium during normal homeostasis and become more active to repair significant wounds. Second, the corneal epithelial stem cell (CESC) hypothesis postulates that, during normal homeostasis, stem cells distributed throughout the basal corneal epithelium, maintain the tissue. According to this hypothesis, LESCs are present in the limbus but are only active during wound healing. We also consider a third possibility, that the corneal epithelium is maintained during normal homeostasis by proliferation of basal corneal epithelial cells without any input from stem cells. After reviewing the published evidence, we conclude that the LESC and CESC hypotheses are consistent with more of the evidence than the third hypothesis, so we do not consider this further. The LESC and CESC hypotheses each have difficulty accounting for one main type of evidence so we evaluate the two key lines of evidence that discriminate between them. Finally, we discuss how lineage-tracing experiments have begun to resolve the debate in favour of the LESC hypothesis. Nevertheless, it also seems likely that some basal corneal epithelial cells can act as long-term progenitors if limbal stem cell function is compromised. Thus, this aspect of the CESC hypothesis may have a lasting impact on our understanding of corneal epithelial maintenance, even if it is eventually shown that stem cells are restricted to the limbus as proposed by the LESC hypothesis.

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“…To prove the existence of markers within our COMECs, putative epithelial stem cell markers, namely ΔP63 α [44, 45] andABCG2 [46, 47], and a specific differentiation marker, cytokeratin K3 (CK3) [14, 48], were firstly examined by RT-PCR. Both of the two groups of COMECs were found to express a corneal and oral epithelial cell marker (CK3) and stem cell markers (ΔNP63 α and ABCG2) (Figures 3 and 4).…”

Section: Resultsmentioning

confidence: 99%

Characterization of Ex Vivo Expanded Oral Mucosal Epithelium Cells on Acellular Porcine Corneal Stroma for Ocular Surface Reconstruction

Wang

Xie

Zhang

2017

Journal of Ophthalmology

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Purpose To ex vivo expand oral mucosal epithelium cells (OMECs) on acellular porcine corneal stroma (APCS) without using feeder cells and serum and to compare the morphologic and phenotypic characteristics of cultured oral cells on APCS to those of cells on deluded human amniotic membrane (HAM). Methods SD rat oral mucosal biopsies were cultured on APCS and HAM. Reverse-transcription polymerase chain reaction (RT-PCR) and immunohistochemistry were used to analyze the characterization of stem cells and epithelial differentiation of the outgrowth products. Results Stratified and optimal transplantable OMECs were obtained after being cultured three to four weeks. Both RT-PCR and immunohistochemistry showed that cultured OMECs expressed markers of epithelial differentiation cytokeratin K3 and epithelial stem cell markers of p63 and ABCG2. Conclusions OMECs can be successfully cultured on APCS without using xenobiotic feeder cells and serum. Characterization showed that these sheets retain the morphologic and phenotypic characteristics of OMECs within differentiated cells and stem cells. The optimal transplantable sheets can prove to be particularly beneficial to both bilateral limbal stem cell deficiency and deep corneal lesions.

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Human limbal epithelium contains side population cells expressing the ATP‐binding cassette transporter ABCG2

Cited by 196 publications

References 40 publications

Ocular surface epithelia contain ABCG2-dependent side population cells exhibiting features associated with stem cells

Ocular surface epithelia contain ABCG2-dependent side population cells exhibiting features associated with stem cells

Evaluating alternative stem cell hypotheses for adult corneal epithelial maintenance

Characterization of Ex Vivo Expanded Oral Mucosal Epithelium Cells on Acellular Porcine Corneal Stroma for Ocular Surface Reconstruction

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