Unknown,Transcriptomics,Genomics,Proteomics

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KSP-positive or -negative cells derived from mouse embryonic stem cells

ABSTRACT: The goals and objectives of this study was to characterize KSP-positive cells derived from mouse embryonic stem cells. KSP is a specific cadherin expressed in renal tubular cells, so it was predicted that KSP-positive cells have characteristics of renal tubular cells. To purify renal lineage cells from differentiated mouse embryonic stem cells, KSP-positive cells were sorted out by use of anti-KSP antibody. Mouse embryonic stem cells were differentiated with Activin 10ng/ml for 18 days through embryoid body formation. 2 independent experiments were carried out. KSP-negative cells were also purified as negative control.

ORGANISM(S): Mus musculus

SUBMITTER: Ryuji Morizane

PROVIDER: E-GEOD-40694 | biostudies-arrayexpress |

REPOSITORIES: biostudies-arrayexpress

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Project description:Purpose: The aim of the present study was to identify and characterize in tissue samples of clear cell-renal cell carcinoma (ccRCC) a population of CD133+/CD24+ cancer cells (RCC-derived cells-RDCs) and to study the differences with their nonneoplastic counterpart, tubular Adult Renal Progenitor Cells (ARPCs). Materials and methods: CD133+/CD24+ RDCs were isolated from 40 patients. The mesenchymal phenotype and stemness proteomic profile of these RDCs were characterized. The colony-forming efficiency and self-renewal ability were tested with limiting dilution. The tumorigenic properties were evaluated in vitro with Soft Agar Assay. The angiogenic response was evaluated in vivo with the chorioallantoic membrane angiogenic assay. Microarray analysis was performed on 6 tARPCs and 6 RDCs clones. Expression of membrane proteins was evaluated with flowcytometry and immunofluorescence staining. Results: CD133+/CD24+ cells were isolated from normal and tumoral kidney tissue. FACS analysis showed that RDCs did not express the mesenchymal stem cell markers. We showed that CD133+/CD24+ tumor cells were more undifferentiated than tARPCs. RDCs were clonigenic and able to differentiate into adipocytes, epithelial and osteogenic cells. RDCs were able to regenerate tumor cells in vitro and to induce angiogenesis in vivo. The gene expression profile identified CTR2 as a membrane marker for this neoplastic population. Conclusions: Our results indicate the presence, in ccRCC, of a CD133+/CD24+/CTR2+ cancer cells population. These cells possess some stem cell-like features, including in vitro self-maintenance and differentiating capabilities, and are able to induce an angiogenic response in vivo. Study of gene expression profiling of Renal Cancer Cells (RCSCs), tubular Adult Renal Progenitor Cells (tARPCs), Renal Proximal Tubular Cells (RPTEC) and Mesenchymal Stem Cells (MSC). To highlight the similarities and the differences with known renal stem populations and with terminally differentiated renal cells, for each patient, 6 subcultures of tubular ARPCs (tARPCs), 3 subcultures of the MSC and 3 subcultures of the RPTEC were included in the analysis.

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KSP-positive or -negative cells derived from mouse embryonic stem cells

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