The in-vitro spheroid culture induces a more highly differentiated but tumorigenic population from melanoma cell lines
| Autor: | Qiang Gu, Baocun Sun, Xiulan Zhao, Ran Sun, Nan Zhao, Jing Mo, Jiadong Chi, Zhiyong Liu, Yuemei Ma, Yanrong Liu, Xueyi Dong |
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| Rok vydání: | 2013 |
| Předmět: |
Cancer Research
Cellular pathology Cellular differentiation Blotting Western Population Fluorescent Antibody Technique Mice Nude Apoptosis Dermatology In Vitro Techniques Biology Real-Time Polymerase Chain Reaction Flow cytometry Immunoenzyme Techniques Mice Cancer stem cell Spheroids Cellular Biomarkers Tumor Tumor Cells Cultured medicine Animals Humans RNA Messenger education Melanoma Cell Proliferation Mice Inbred BALB C education.field_of_study medicine.diagnostic_test Reverse Transcriptase Polymerase Chain Reaction ABCB5 Cell Differentiation Flow Cytometry medicine.disease Oncology Neoplastic Stem Cells Cancer research Heterografts Female Stem cell |
| Zdroj: | Melanoma Research. 23:254-263 |
| ISSN: | 0960-8931 |
| Popis: | Cancer stem cells (CSCs) have been identified in various malignancies, and different properties have been examined to characterize CSCs: tumorigenicity in immunocompromised mice, stem cell surface markers, label-retaining properties, and proliferation as nonadherent spheres. This study explored the consistency and efficiency among these methods. Among the melanoma cell lines examined (A375, A875, MUM-2b, and MUM-2c), only A375 and MUM-2c grew as nonadherent spheres and continuously propagated in a defined serum-free medium in vitro. Flow cytometry and immunofluorescence analysis indicated that sphere-derived cells contained a smaller proportion of cells expressing the candidate surface markers of melanoma stem cells such as ABCB5, CD133, CD20 and CD271, and a larger proportion of cells expressing melanocytic differentiation markers such as HMB45 and S100 protein, compared with adherent cells. Surprisingly, the more highly differentiated sphere-derived melanoma cells exhibited increased tumorigenic potential in vivo, as indicated by shorter tumor incubation (A375) and smaller number of cells required to initiate tumor formation (A375 and MUM-2c) compared with those of parental cells. Despite the similarity in histopathological characteristics, the expression profile indicated that xenografts derived from sphere-derived melanoma cells exhibited a more tumorigenic phenotype with respect to the stem or the differentiation markers detected by immunohistochemical analysis. Therefore, sphere formation in nonadherent cultures may not be a preferred surrogate in-vitro method for enriching melanoma stem cells according to candidate markers but may be a favorable condition for activating potential CSCs. |
| Databáze: | OpenAIRE |
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