Project description:PCL family protein Phf19/Pcl3 is one of the accessory components of the PRC2 core complex, and Phf19 is highly expressed in murine ES cells and an ES cell-like embryonic carcinoma cell line, F9 cells. Here we performed microarray analysis of embryonal carcinoma cell line F9 following Phf19 knockdown by shRNA. Knocking down Phf19/Pcl3 in F9 embryonic cells led to derepression of numerous PRC2 direct target genes.
Project description:PCL family protein Phf19/Pcl3 is one of the accessory components of the PRC2 core complex, and Phf19 is highly expressed in murine ES cells and an ES cell-like embryonic carcinoma cell line, F9 cells. Here we performed microarray analysis of embryonal carcinoma cell line F9 following Phf19 knockdown by shRNA. Knocking down Phf19/Pcl3 in F9 embryonic cells led to derepression of numerous PRC2 direct target genes. 4 sampels including 2 shRNA vector control cell lines and 2 shPhf19 cell lines were used for RNA extraction and Affymetrix mouse 430 2.0 arrays.
Project description:This SuperSeries is composed of the following subset Series: GSE30537: Dissecting the retinoid-induced differentiation of F9 embryonal stem cells by integrative genomics [mRNA profiling] GSE30538: Dissecting the retinoid-induced differentiation of F9 embryonal stem cells by integrative genomics [ChIP-seq] Refer to individual Series
Project description:Here we perfomed the Teratoma assay for a normal human embryonic stem cell line (H9(+Dox)), a human embryonic stem cell line with a mesendodermal differentiation bias (H9Hyb), a normal human induced pluripotent stem cell line (LU07), a human induced pluripotent stem cell line with reactivated transgenes (LU07+Dox) and a human embryonal carcinoma cell line (EC) and anayzed their gene expression. The ability to form teratomas in vivo containing multiple somatic cell types is regarded as functional evidence of pluripotency for human pluripotent stem cells (hPSCs). Since the Teratoma assay is animal-dependent, laborious and only qualitative, the PluriTest and the hPSC ScoreCard assay have been developed as in vitro alternatives. Here we compared normal hPSCs, induced hPSCs (hiPSCs) with reactivated reprogramming transgenes and human embryonal carcinoma (hEC) cells in these assays. While normal hPSCs gave rise to typical teratomas, the xenografts of the hEC cells and the hiPSCs with reactivated reprogramming transgenes were largely undifferentiated and malignant. The hPSC ScoreCard assay confirmed the line-specific differentiation propensities in vitro. However, when undifferentiated cells were analysed with PluriTest only hEC cells were identified as abnormal whereas all other cell lines were indistinguishable and resembled normal hPSCs. Our results indicate that pluripotency assays are best selected on the basis of intended downstream applications.
Project description:Here we analyzed undifferentiated cells and perfomed the Teratoma assay for a normal human embryonic stem cell line (H9(+Dox)), a human embryonic stem cell line with a mesendodermal differentiation bias (H9Hyb), a normal human induced pluripotent stem cell line (LU07), a human induced pluripotent stem cell line with reactivated transgenes (LU07+Dox) and a human embryonal carcinoma cell line (EC). The ability to form teratomas in vivo containing multiple somatic cell types is regarded as functional evidence of pluripotency for human pluripotent stem cells (hPSCs). Since the Teratoma assay is animal-dependent, laborious and only qualitative, the PluriTest and the hPSC ScoreCard assay have been developed as in vitro alternatives. Here we compared normal hPSCs, induced hPSCs (hiPSCs) with reactivated reprogramming transgenes and human embryonal carcinoma (hEC) cells in these assays. While normal hPSCs gave rise to typical teratomas, the xenografts of the hEC cells and the hiPSCs with reactivated reprogramming transgenes were largely undifferentiated and malignant. The hPSC ScoreCard assay confirmed the line-specific differentiation propensities in vitro. However, when undifferentiated cells were analysed with PluriTest only hEC cells were identified as abnormal whereas all other cell lines were indistinguishable and resembled normal hPSCs. Our results indicate that pluripotency assays are best selected on the basis of intended downstream applications.
Project description:SOX2 is an oncogene and a core pluripotency transcription factor. SOX2 has multiple roles in various malignancies, in the maintainance of pluripotency and during various stages of embryonic development. Human embryonal carcinoma cells express SOX2 and the loss of this results in their differentiation. We silenced SOX2 in two human embryonal carcinoma cell lines and measured whole-genome mRNA expression. Many genes related to embryogenesis and tissue morphogensis were upregulated. Other upregulated genes were markers of mesodermal development and epithelial-to-mesenchymal transition. A specific and validated siRNA against SOX2 was chemically transfected in undifferentiated 2102Ep and NTera-2 embryonal carcinoma cell lines. After three days of incubation under normal growth conditions we used Affymetrix microarrays to measure whole-genome mRNA transcript expression in three biological replicates of each cell line and compared this to whole-gene expression in identical samples transfected with a non-targeting, scrambled control siRNA. SOX2 silencing was validated using qRT-PCR and Western blot prior to whole-genome expression analysis.
Project description:Gene expression profiles of undifferentiated mouse embryonal carcinoma cell strains (P19, P19CL6, and 4 P19CL6 sublines) were obtained, using Affymetrix GeneChip Mouse Genome 430A and 430B. Heart diseases such as cardiac infarction damage cardiomyocytes and consequently lead to significant loss of the contractile capacity of the heart. To repair functions of the injured heart, a great deal of research has attempted to develop regenerative medicine using pluripotent stem cell-based cardiomyocytes as cell therapy products. However, the efficiency of the current methods available for the cardiac differentiation of stem cells is insufficient for clinical settings. A comprehensive understanding of the mechanism involved in the cardiac differentiation of stem cells is necessary to improve the differentiation efficiency. To identify genes assosiated with cardiomyogenic potential, we isolated P19CL6 cell sublines possessing distinct properties in cardiomyogenesis and comprared their transcriptome profiles with those of mouse embryonal carcinoma P19 and P19CL6 cells. Total RNA isolated from undifferentiated EC cell strains (P19 cells, P19CL6 cells, and 4 P19CL6 sublines) using Affymetrix chips MOE430A and MOE430AB. CEL files unavailable.
Project description:Gene expression profiles of undifferentiated mouse embryonal carcinoma cell strains (P19, P19CL6, and 4 P19CL6 sublines) were obtained, using Affymetrix GeneChip Mouse Genome 430A and 430B. Heart diseases such as cardiac infarction damage cardiomyocytes and consequently lead to significant loss of the contractile capacity of the heart. To repair functions of the injured heart, a great deal of research has attempted to develop regenerative medicine using pluripotent stem cell-based cardiomyocytes as cell therapy products. However, the efficiency of the current methods available for the cardiac differentiation of stem cells is insufficient for clinical settings. A comprehensive understanding of the mechanism involved in the cardiac differentiation of stem cells is necessary to improve the differentiation efficiency. To identify genes assosiated with cardiomyogenic potential, we isolated P19CL6 cell sublines possessing distinct properties in cardiomyogenesis and comprared their transcriptome profiles with those of mouse embryonal carcinoma P19 and P19CL6 cells.