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Archive > Volume 21 Issue 3 > 2014,21(3):245-250. DOI:10.3872/j.issn.1007-385X.2014.03.002 Prev Next
Prompt Report
Mesenchymal differentiation potential of single-cell cloned liver cancer stem cells
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Abstract:
To investigate the multilineage differentiation potential of single-cell-cloned liver cancer stem cells (LCSCs). Methods: Single cell cloned LCSCs were generated by limiting dilution cloning and confirmed by RT-PCR analysis of stem cell markers. Phenotype-confirmed LCSCs were induced to differentiate into osteoblasts, chondrocytes and adipocytes, respectively, by culturing them in corresponding differentiation-inducing media for 3 weeks. Both before and after the differentiation induction culture, the expression of markers specific for osteoblasts, chondrocytes and adipocytes was comparatively analyzed by real-time PCR and cell type-specific staining techniques.Results: Stem cell markers including CD133, CD34, ATP-binding cassette sub-family G member 2 (ABCG2), nestin, stem cell factor (SCF) and stem cell growth factor receptor (C-kit) were detected in undifferentiated single cell cloned LCSCs. After three weeks of differentiation induction, cells cultured in the osteoblast-specific medium formed orange calcium nodules as demonstrated by Alizarin red staining; cells cultured in the chondrocyte-specific medium showed blue proteoglycan depositions as revealed by Alcian blue staining; and cells cultured in the adipocyte-specific medium showed Oil Red O stained lipid droplets. In the three corresponding differentiation induction cultures, LCSCs acquired the expression of markers for osteoblast (osteocalcin and collagen), chondrocyte (aggrecan and collagen type Ⅱ), and adipocyte (adiponectin and peroxisome proliferator-activated receptor) respectively. All the differences were significant (P<0.01) except for collagen type Ⅰand collagen type Ⅱ (P<0.05). Conclusion: These preliminary observations suggest that single cell cloned LCSCs possess a plasticity and may potentially differentiate into mesenchymal-like cells under specific microenvironments.
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Project Supported:
Project supported by the Major National Basic Research Development Program (973 program) of China (No. 2009CB521804), and the National Natural Science Foundation of China (No. 81370873, No. 81302334)
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