Isolation of Cancer Stem Cells from Squamous Cell Carcinoma

Fontenete, Sílvia; Pérez-Moreno, Mirna

doi:10.1007/7651_2018_162

Cited by 6 publications

(3 citation statements)

References 13 publications

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“…Different mouse models of SCC and BCC are widely used for studying stem cell, cancer stem cell, and cancer biology. 57 , 58 , 59 , 60 , 61 , 62 , 63 , 64 , 65 , 66 To determine the relevance of established mouse models to human skin cancer, we first determined whether the mouse SCC stem cell (SCC-SC) signature genes are highly represented in the human SCC. 66 We found that 598 mouse SCC-SC genes with human orthologs were expressed in our human tumor or normal tissues.…”

Section: Resultsmentioning

confidence: 99%

Distinct transcriptomic landscapes of cutaneous basal cell carcinomas and squamous cell carcinomas

et al. 2021

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The majority of non-melanoma skin cancer (NMSC) is cutaneous basal cell carcinoma (BCC) or squamous cell carcinoma (SCC), which are also called keratinocyte carcinomas, as both of them originate from keratinocytes. The incidence of keratinocyte carcinomas is over 5 million per year in the US, three-fold higher than the total incidence of all other types of cancer combined. While there are several reports on gene expression profiling of BCC and SCC, there are significant variations in the reported gene expression changes in different studies. One reason is that tumor-adjacent normal skin specimens were not included in many studies as matched controls. Furthermore, while numerous studies of skin stem cells in mouse models have been reported, their relevance to human skin cancer remains unknown. In this report, we analyzed gene expression profiles of paired specimens of keratinocyte carcinomas with their matched normal skin tissues as the control. Among several novel findings, we discovered a significant number of zinc finger encoding genes up-regulated in human BCC. In BCC, a novel link was found between hedgehog signaling, Wnt signaling, and the cilium. While the SCC cancer-stem-cell gene signature is shared between human and mouse SCCs, the hair follicle stem-cell signature of mice was not highly represented in human SCC. Differential gene expression (DEG) in human BCC shares gene signature with both bulge and epidermal stem cells. We have also determined that human BCCs and SCCs have distinct gene expression patterns, and some of them are not fully reflected in current mouse models.

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Section: Resultsmentioning

confidence: 99%

Distinct transcriptomic landscapes of cutaneous basal cell carcinomas and squamous cell carcinomas

et al. 2021

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“…SCCs or skin samples from prior SCC locations (n = 42) were dissected and processed as described by Fontenete et al [26]. Single-cell suspensions were stained using different antibodies: CD45-FITC (BD Pharmingen, San Diego, CA), CD3-APC (eBioscience, San Diego, CA), Cd11b-PerCPCy5.5 (eBioscience), B220-PE-Cy7 (eBioscience), Ly6G-PE (Biolegend, San Diego, CA), CD45-FITC (BD Pharmingen), CD8-PE-Cyanine7 (Biolegend), and CD3-APC (eBioscience).…”

Section: Flow Cytometrymentioning

confidence: 99%

Tumor Clearance and Immune Cell Recruitment in UV‐Induced Murine Squamous Cell Carcinoma Exposed to Ablative Fractional Laser and Imiquimod Treatment

et al. 2021

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Background and Objectives: Keratinocyte carcinoma (KC) is the most common cancer worldwide, and squamous cell carcinoma (SCC) is the second most frequent subtype. Ablative fractional laser (AFL)-assisted drug delivery significantly enhances the uptake of topically applied drugs. The objective of this study was to assess tumor response and perform a descriptive characterization of the local recruitment of immune cells and systemic immune mediator levels in an ultraviolet radiation (UVR)-induced murine SCC model after AFL treatment alone and combined with topical imiquimod. Study Design/Materials and Methods: Immunocompetent hairless mice (C3•Cg/TifBomTac, n = 74) were irradiated with solar-simulated UVR until 3-mm SCCs developed. The mice were divided into four interventional groups: AFL alone, AFL + imiquimod, imiquimod alone, and untreated SCC controls. AFL was given as a single treatment, whereas imiquimod was applied daily until the mice were euthanized on Days 0, 2, 7, or 14. SCCs were photographed and measured (mm) to assess the therapeutic response. Skin samples were processed for histopathological and immunohistochemical analyses, as well as for flow cytometry. Cytokine expression changes in sera were analyzed using ELISpot cytokine arrays. Results: Treatment of mouse SCCs with AFL + imiquimod induced the most robust immune cell infiltration and the greatest proportion of tumor clearance compared to other interventions. Early innate immune cell infiltration was induced by AFL + imiquimod treatment as the number of neutrophils and macrophages had increased fourfold within 2 days of treatment initiation compared with untreated SCC control mice (P < 0.05). AFL treatment alone had a more limited effect, with a fourfold increase in neutrophils (P < 0.05) but no significant increase in the number of macrophages. Correspondingly, treatment with AFL + imiquimod had the greatest effects on the adaptive immune cell recruitment: CD4 + T-helper cells increased threefold at Day 7 compared with untreated SCCs (P = 0.0001) and, notably, cytotoxic CD8 + T cells increased 14-fold at Day 14 (P = 0.0112). In addition, FOXP3 + regulatory T cells (Tregs) increased 14-fold at Day 7 (P = 0.0026), suggesting the resolution of the inflammatory infiltration. AFL treatment alone induced a moderate immune cell infiltration (a twofold increase in CD4 + T-helper cells, P = 0.0200; a threefold increase in CD8 + T cells, P = 0.0100; and a 14-fold increase in FOXP3 + Tregs at Day 14, P = 0.0021), whereas imiquimod alone did not significantly increase cell counts. AFL + imiquimod treatment increased CXCL12 serum levels threefold at Day 14 (P = 0.0200). Conclusion: AFL treatment alone and in combination with imiquimod induces substantial tumor clearance associated with local recruitment of innate and adaptive immune cells in UVR-induced murine SCCs. These results may provide a basis for new immunotherapeutic approaches to KC treatment.

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“…HF-MSCs CD34 may be to refresh hematopoietic system, as mentioned previously (Lako et al 2002 ). Some studies were focused on the surface marker of HF-MSCs such as integrin, keratin (CK14, CK15, and CK19), C8/144B, p63, CD71 and CX43 (Dong et al 2014 ; Maleki et al 2014 ; Son et al 2015 ), CD34 (Sol et al 2018 ) CD49f (Krebsbach and Villa-Diaz 2017 ) and CD34 and CD49f duel-positive cell residents in mice (Fontenete and Perez-Moreno 2018 ). Babakhani et al (Babakhani et al 2019 ) showed that the bulge of HF-MSCs was nestin and CD34 positive and Kr15 negative.…”

Section: Discussionmentioning

confidence: 99%

Comparative characterization and osteogenic / adipogenic differentiation of mesenchymal stem cells derived from male rat hair follicles and bone marrow

2020

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Clinical applications of cell therapy and tissue regeneration under different conditions need a multiplicity of adult stem cell sources. Up to date, little is available on the comparative isolation, characterization, proliferation, rapid amplification, and osteogenic/adipogenic differentiation of rat mesenchymal stem cells (MSCs) isolated from living bulge cells of the hair follicle (HF) and bone marrow (BM) from the same animal. This work hopes to use HF-MSCs as an additional adult stem cell source for research and application. After reaching 80% confluence, the cell counting, viability %, and yields of HF-MSCs and BM-MSCs were nearly similar. The viability % was 91.41 ± 2.98 and 93.11 ± 3.06 while the cells yield of initial seeding was 33.15 ± 2.76 and 34.22 ± 3.99 and of second passage was 28.76 ± 1.01 and 29.56 ± 3.11 for HF-MSCs and BM-MSCs respectively. Clusters of differentiation (CDs) analysis revealed that HF-MSCs were positively expressed CD34, CD73 and CD200 and negatively expressed CD45. BM-MSCs were positively expressed CD73 and CD200 and negatively expressed of CD34 and CD45. The proliferation of HF-MSCs and BM-MSCs was determined by means of incorporation of Brd-U, population doubling time (PDT) assays and the quantity of formazan release. The percentage of Brd-U positive cells and PDT were relatively similar in both types of cells. The proliferation, as expressed by the quantity of formazan assay in confluent cells, revealed that the quantity of release by BM-MSCs was slightly higher than HF-MSCs. Adipogenic differentiated BM-MSCs showed moderate accumulation of oil red-O stained lipid droplets when compared to that of HF-MSCs which exhibited high stain. The total lipid concentration was significantly higher in adipogenic differentiated HF-MSCs than BM-MSCs (P < 0.05). It was found that activity of bone alkaline phosphatase and calcium concentration were significantly higher (P < 0.01 and P < 0.05 respectively) in osteogenic differentiated BM-MSCs than that of HF-MSCs. The present findings demonstrate that the HF-MSCs are very similar in most tested characteristics to BM-MSCs with the exception of differentiation. Additionally; no issues have been reported during the collection of HF-MSCs. Therefore, the HF may represent a suitable and accessible source for adult stem cells and can be considered an ideal cell source for adipogenesis research.

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Isolation of Cancer Stem Cells from Squamous Cell Carcinoma

Cited by 6 publications

References 13 publications

Distinct transcriptomic landscapes of cutaneous basal cell carcinomas and squamous cell carcinomas

Distinct transcriptomic landscapes of cutaneous basal cell carcinomas and squamous cell carcinomas

Tumor Clearance and Immune Cell Recruitment in UV‐Induced Murine Squamous Cell Carcinoma Exposed to Ablative Fractional Laser and Imiquimod Treatment

Comparative characterization and osteogenic / adipogenic differentiation of mesenchymal stem cells derived from male rat hair follicles and bone marrow

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