Metabolomics,Unknown,Transcriptomics,Genomics,Proteomics

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Single-cell RNA-seq (10x Chromium V3) of heterogeneous spheres consisting of melanoma cell line and two types of human primary dermal fibroblasts

ABSTRACT: Photo-damage represents an important aspect of cutaneous carcinogenesis. The photo-damaged microenvironment significantly facilitates progression of malignant melanoma. Therefore human primary dermal fibroblasts from photo-damaged adult skin and juvenile sun-protected skin were isolated and expanded as described by Dvorankova et al. (PMID: 29675784). Low passage fibroblasts were mixed in suspension (in ratio 1:1) with G361 malignant melanoma cell line (CVCL_1220). Using the hanging drop technique (PMID: 29786551), the melanoma/fibroblast spheres containing 50,000 cells per sphere were formed during the next 72 hours. The organoids were consequently maintained for another 2 days in non-adhesive culture wells in complete DMEM culture medium. At this time, heterogeneous spheres were harvested for immunohistochemical analysis or for further invasion studies. Single-cell suspensions from heterogeneous spheres were analysed by single-cell RNA sequencing (10x Chromium V3) to reveal the transcriptional heterogeneity of model cell populations under given conditions with emphasis on the features of dermal fibroblasts exposed to the influence of malignant melanoma.

INSTRUMENT(S): NextSeq 500

ORGANISM(S): Homo sapiens

SUBMITTER: Jiri Novotny 

PROVIDER: E-MTAB-9216 | biostudies-arrayexpress |

REPOSITORIES: biostudies-arrayexpress

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Publications

Analysis of HPV-Positive and HPV-Negative Head and Neck Squamous Cell Carcinomas and Paired Normal Mucosae Reveals Cyclin D1 Deregulation and Compensatory Effect of Cyclin D2.

Novotný Jiří J   Bandúrová Veronika V   Strnad Hynek H   Chovanec Martin M   Hradilová Miluše M   Šáchová Jana J   Šteffl Martin M   Grušanović Josipa J   Kodet Roman R   Pačes Václav V   Lacina Lukáš L   Smetana Karel K   Plzák Jan J   Kolář Michal M   Vomastek Tomáš T  

Cancers 20200326 4

Aberrant regulation of the cell cycle is a typical feature of all forms of cancer. In head and neck squamous cell carcinoma (HNSCC), it is often associated with the overexpression of cyclin D1 (<i>CCND1</i>). However, it remains unclear how <i>CCND1</i> expression changes between tumor and normal tissues and whether human papillomavirus (HPV) affects differential <i>CCND1</i> expression. Here, we evaluated the expression of D-type cyclins in a cohort of 94 HNSCC patients of which 82 were subject  ...[more]

PMID: 32224897

Publication: 1/2

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