Identification of Epithelial Stem Cells In Vivo and In Vitro Using Keratin 19 and BrdU

Larouche, Danielle; Lavoie, Amélie; Paquet, Claudie; Simard-Bisson, Carolyne; Germain, Lucie

doi:10.1007/978-1-60761-380-0_27

Cited by 19 publications

(9 citation statements)

References 5 publications

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“…The lack of immunoreactivity demonstrated that these keratinocytes actually lacked the CK19 antigen; CK19 is considered to be a putative stem cell marker, and it is only expressed in the most basal layer, which might relate to the local environments that penetrate the basement membrane (Larouche et al, 2005;Larouche et al, 2010;Michel et al, 1996).…”

Section: Accepted Manuscriptmentioning

confidence: 95%

Histological features of oral epithelium in seven animal species: As a reference for selecting animal models

Xiong

et al. 2016

European Journal of Pharmaceutical Sciences

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Section: Accepted Manuscriptmentioning

confidence: 95%

Histological features of oral epithelium in seven animal species: As a reference for selecting animal models

Xiong

et al. 2016

European Journal of Pharmaceutical Sciences

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“…Nevertheless, both studies showed the presence of CK19-positive cells in MCC [59, 62]. CK19 has been used as an epidermal stem cell marker [64], but is also present on mature Merkel cells [65]. Moreover, Abbas and Bhawan found no CK15-positive cells in MCC [62].…”

Section: Putative Cells Of Origin For MCCmentioning

confidence: 99%

Which Are the Cells of Origin in Merkel Cell Carcinoma?

Tilling

Moll

2012

Journal of Skin Cancer

119

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Merkel cell carcinoma (MCC), a highly aggressive skin tumour with increasing incidence, is associated with the newly discovered Merkel cell polyomavirus (MCPyV). Studies on MCC and MCPyV as well as other risk factors have significantly increased our knowledge of MCC pathogenesis, but the cells of origin, which could be important targets in future therapies, are still unknown. Merkel cells (MCs), the neuroendocrine cells of the skin, were believed to be at the origin of MCC due to their phenotypic similarities. However, for several reasons, for example, heterogeneous differentiation of MCCs and postmitotic character of MCs, it is not very likely that MCC develops from differentiated MCs. Skin stem cells, probably from the epidermal lineage, are more likely to be cells of origin in MCC. Future studies will have to address these questions more directly in order to identify the physiological cells which are transformed to MCC cells.

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“…Proliferation rate was examined with 5-bromodeoxyuridine (BrdU)-pulsed cultures as previously described [32]. Briefly, melanoma cells were plated onto AcOH or chitosan in culture medium containing 10 µM BrdU for 24h.…”

Section: Methodsmentioning

confidence: 99%

Anticancer properties of chitosan on human melanoma are cell line dependent

Gibot

Chabaud

Bouhout

et al. 2015

International Journal of Biological Macromolecules

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Purpose: Chitosan, a natural macromolecule, is widely used in medical and pharmaceutical fields because of its distinctive properties such as bactericide, fungicide and above all its antitumor effects. Although its antitumor activity against different types of cancer had been previously described, its mechanism of action was not fully understood. Materials and Methods: Coating of chitosan has been used in cell cultures with A375, SKMEL28 and RPMI7951 cell lines. Adherence, proliferation and apoptosis were investigated. Results: Our results revealed that whereas chitosan decreased adhesion of primary melanoma A375 cell line and decreased proliferation of primary melanoma SKMEL28 cell line, it had potent pro-apoptotic effects against RPMI7951, a metastatic melanoma cell line. In these latter cells, inhibition of specific caspases confirmed that apoptosis was effected through the mitochondrial pathway and Western blot analyses showed that chitosan induced an up regulation of pro-apoptotic molecules such as Bax and a down regulation of anti-apoptotic proteins like Bcl-2 and Bcl-XL. More interestingly, chitosan exposure induced an exposition of a greater number of CD95 receptor at RPMI7951 surface, making them more susceptible to FasL-induced apoptosis. Conclusion: Our results indicate that chitosan could be a promising agent for further evaluations in antitumor treatments targeting melanoma. Point 1: This point is very interesting, notably to verify if lower doses could also affect melanoma cells and especially the RPMI7951 metastatic cancer cells. It could also be pertinent to minimize adverse effect on healthy cells. We have evaluated the effect of chitosan concentration on RPMI7951 cell line and normal primary human dermal fibroblasts and it is reported on Fig.5C (for caspase-3 activity) and D (for cell count using WST-1 test).Point 2: Previous the first submission, we had done tests to verify whether apoptosis induction was detectable on fibroblasts but not extensively. Thus, the remark of the reviewer was a chance to improve this paper. Chitosan effect was assessed on adhesion, proliferation and apoptosis of normal primary human dermal fibroblasts. As depicted in fig.5A, apoptotic morphology was not seen when fibroblasts were cultured on chitosan coating as well as no/low caspase activation (Fig.5C, even at high doses of Chitosan, and Fig.5F, even when cultures were continued for 6 days). However, it could be noted that the cell number was reduced ( Fig.5D and E), probably as a result of a reduced proliferation. In vivo, because fibroblasts, unlike cancer cells, have a very slow proliferation rate, chitosan should have a low action on these cells. Cell attachment was tested (Fig.5B) and, even if a delay could be observed in the presence of chitosan after 6 hours, virtually all fibroblasts had adhered on the plate. Point 3: Long term cultures (here 6 days) have been tested. Caspase activation increased substantially when RPMI7951 were seeded onto chitosan coated plates but such a phenomenon could not b...

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Identification of Epithelial Stem Cells In Vivo and In Vitro Using Keratin 19 and BrdU

Cited by 19 publications

References 5 publications

Histological features of oral epithelium in seven animal species: As a reference for selecting animal models

Histological features of oral epithelium in seven animal species: As a reference for selecting animal models

Which Are the Cells of Origin in Merkel Cell Carcinoma?

Anticancer properties of chitosan on human melanoma are cell line dependent

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