Comparable mRNA expression of pluripotency-related genes in SNUhES3 cells cultured on autofeeder or xenofeeder system
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ABSTRACT:
ORGANISM(S): Homo sapiens
PROVIDER: GSE26993 | GEO | 2011/02/02
SECONDARY ACCESSION(S): PRJNA136933
REPOSITORIES: GEO
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Project description:I developed a new culture system for hES cells; this system does not require supplementation with bFGF to obtain hES cells that are suitable for tissue engineering and regenerative medicine. This culture system employed mesenchymal stem cells derived from hES cells (hESC-MSCs) as autologous human feeder cells in the absence of bFGF. For pluripotency-related gene expression profiling, a cDNA microarray analysis was performed SNUhES3 cultured on the autofeeder hESC-MSCs layer maintained the undifferentiated state for 30 passages in a manner similar to the SNUhES3 cells on xenofeeder STO cell layer. To compare the pluripotency-related genes in SNUhES3 cells cultured on autofeeder or xenofeeder system, I used agilent one-color array. Two independant experiment performed.
2011-02-02 | E-GEOD-26993 | biostudies-arrayexpress
Project description:To assess the effect of sodium butyrate exposure on human ESC grown without culture support for self-renewal (I.e. without conditioned medium and added bFGF) in support of data generated on H1 hESC - two groups were compared - BG02 culture in feeder conditioned versus on sodium butyrate - in triplicate and compared on Agilent whole human genome array BG02 hESC were grown under two conitions - A. for 31 passages on Matrigel in feeder conditioned medium and B. for 29 passages on Matrigel in feeder conditioned medium followed by 20 passages in 0.2 mM sodium butyrate without conditioned medium and in human ES cell medium containing no added bFGF
2009-03-04 | E-GEOD-15109 | biostudies-arrayexpress
Project description:To assess the effect of sodium butyrate exposure on human ESC grown without culture support for self-renewal (I.e. without conditioned medium and added bFGF) in support of data generated on H1 hESC - two groups were compared - BG02 culture in feeder conditioned versus on sodium butyrate - in triplicate and compared on Agilent whole human genome array
2009-03-05 | GSE15109 | GEO
Project description:Human embryonic stem (hES) cells have the capacities to propagate for extended periods and to differentiate into cell types from all three germ layers both in vitro and in vivo. These characteristics of self-renewal and pluripotency enable hES cells having the potential to provide an unlimited supply of different cell types for tissue replacement, drug screening, and functional genomics studies. The hES-T3 cells with normal female karyotype cultured on either mouse embryonic fibroblasts (MEF) in hES medium (containing 4 ng/ml bFGF) (T3MF) or feeder-free Matrigel in MEF-conditioned medium (supplemented with additional 4 ng/ml bFGF) (T3CM) were found to express very similar profiles of mRNAs and microRNAs, indicating that the unlimited self-renewal and pluripotency of hES cells can be maintained by continuing culture on these two conditions. However, the expression profiles, especially microRNAs, of the hES-T3 cells cultured on Matrigel in hES medium supplemented with 4 ng/ml bFGF and 5 ng/ml activin A (T3BA) were found to be different from those of T3MF and T3CM cells. In T3BA cells, four hES cell-specific microRNAs miR-372, miR-302d, miR-367 and miR-200c, as well as three other microRNAs miR-199a, miR-19a and miR-217, were found to be up-regulated, whereas five miRNAs miR-19b, miR-221, miR-222, let-7b and let-7c were down-regulated by activin A. Thirteen abundantly differentially expressed mRNAs, including NR4A2, ERBB4, CXCR4, PCDH9, TMEFF2, CD24 and COX6A1 genes, targeted by seven over-expressed miRNAs were identified by inverse expression levels of these seven microRNAs to their target mRNAs in T3BA and T3CM cells. The NR4A2, ERBB4 and CXCR4 target genes were further found to be regulated by EGF and/or TNF. The 50 abundantly differentially expressed genes targeted by five under-expressed miRNAs were also identified. The abundantly expressed mRNAs in T3BA and T3CM cells were also analyzed for the network and signaling pathways, and roles of activin A in cell proliferation and differentiation were found. These findings will help elucidate the complex signaling network which maintains the self-renewal and pluripotency of hES cells. In this investigation, both miRNA and mRNA expression profiles from human embryonic stem cells grown on MEF feeder (T3MF), feeder-free Matrigel in MEF-conditioned medium (T3CM) and in hES medium (containing 4 ng/ml bFGF) supplemented with 5 ng/ml activin A (T3BA) were quantitatively determined. Several target genes of T3BA and T3CM cells-specific miRNAs were identified. ***This submission represents the mRNA expression component of the study only***
2010-09-10 | E-GEOD-16910 | biostudies-arrayexpress
Project description:To assess the effect of sodium butyrate exposure on human ESC grown without culture support for self-renewal (I.e. without conditioned medium and added bFGF) - three groups were compared - H1 culture in feeder conditioned medium vs without conditioned medium in 0.2 mM sodium butyrate vs. grown in sodium butyrate for 4 passages followed by return to conditioned medium conditions for 3 passages. The three groups were grown in triplicate and compared on Agilent whole human genome array Experiment Overall Design: H1 hESC were grown under 3 conditions: A. 48 passages total, the last 9 passages off of feeders on Matrigel in feeder conditioned medium B. 48 passages total, within the last 9 passges, #39-41 were in conditioned medium and #42-48 were without conditioned medium and added bFGF but with 0.2 mM sodium butyrate on Matrigel C. 49 passages total, within the last 10 passages #39-41 were in conditioned medium, #42-46 in 0.2 mM sodium butyrate (without feeder of bFGF support) and #47-49 back to conditioned medium + bFGF. The RNA from these cells were compared via Agilent whole human genome array.
2009-03-15 | E-GEOD-15110 | biostudies-arrayexpress
Project description:Analysis of KhES-1 and H9 human ES cells in growth factors-dependent (E8) and -independent (AKIT) medium in feeder-free culture condition and KSR/bFGF medium on a feeder-layer. Results provide insight into genetic stability in different culture media/conditions.
2017-09-08 | GSE103636 | GEO
Project description:Human embryonic stem (hES) cells have the capacities to propagate for extended periods and to differentiate into cell types from all three germ layers both in vitro and in vivo. These characteristics of self-renewal and pluripotency enable hES cells having the potential to provide an unlimited supply of different cell types for tissue replacement, drug screening, and functional genomics studies. The hES-T3 cells with normal female karyotype cultured on either mouse embryonic fibroblasts (MEF) in hES medium (containing 4 ng/ml bFGF) (T3MF) or feeder-free Matrigel in MEF-conditioned medium (supplemented with additional 4 ng/ml bFGF) (T3CM) were found to express very similar profiles of mRNAs and microRNAs, indicating that the unlimited self-renewal and pluripotency of hES cells can be maintained by continuing culture on these two conditions. However, the expression profiles, especially microRNAs, of the hES-T3 cells cultured on Matrigel in hES medium supplemented with 4 ng/ml bFGF and 5 ng/ml activin A (T3BA) were found to be different from those of T3MF and T3CM cells. In T3BA cells, four hES cell-specific microRNAs miR-372, miR-302d, miR-367 and miR-200c, as well as three other microRNAs miR-199a, miR-19a and miR-217, were found to be up-regulated, whereas five miRNAs miR-19b, miR-221, miR-222, let-7b and let-7c were down-regulated by activin A. Thirteen abundantly differentially expressed mRNAs, including NR4A2, ERBB4, CXCR4, PCDH9, TMEFF2, CD24 and COX6A1 genes, targeted by seven over-expressed miRNAs were identified by inverse expression levels of these seven microRNAs to their target mRNAs in T3BA and T3CM cells. The NR4A2, ERBB4 and CXCR4 target genes were further found to be regulated by EGF and/or TNF. The 50 abundantly differentially expressed genes targeted by five under-expressed miRNAs were also identified. The abundantly expressed mRNAs in T3BA and T3CM cells were also analyzed for the network and signaling pathways, and roles of activin A in cell proliferation and differentiation were found. These findings will help elucidate the complex signaling network which maintains the self-renewal and pluripotency of hES cells.
2010-09-10 | GSE16910 | GEO
Project description:Purpose: 1. Compare transcriptomes of hESC-derived MSCs (EMSCs) and somatic MSCs (SMCS); 2. Reveal transriptomic profiles of EMSCs post injection into mouse blastocyst. Methods: For bulk seq on cultured human MSCs, mRNA was isolated using RNeasy Mini Kit (QIAGEN) and libraries was constructed using an Illumina kit, and then sequenced on NovaSeq platform. For single cell (sc) RNAseq on cultured human MSCs, cells were dissociated and lysed in the Oil-droplet Single Cell Capture System (Nadia Dolomite) and libraries were constructed using Nextera kits (Illumina), and then sequenced on Hiseq 150PE (Illumina). For EMSCs injected into mouse blastocyst, 30~70 embryos were pulled and lyzed in 10uL of Lysis Buffer for SmartSeq. Results: 1. Both in bulk and in single cell level, all types of MSCs share typical common markers; 2. Unignorable differences exhist among hES-derived and somatic MSCs; 3. Rapid mRNA profile shifts in metabolism, cellular skeleton, and immunoregulatory response happen in hES-derived MSCs post injection to mouse embryos. Conclusions: These experiments reveal transcriptomic alterations of EMSCs after injection into mouse blastocyst and transcriptomic profiles of human MSCs from different sources.
2023-11-21 | GSE195573 | GEO
Project description:Background: The hES-T3 cell line with normal female karyotype was used to differentiate into autogeneic fibroblast-like cells (T3HDF) as feeder to support the undifferentiated growth of hES-T3 cells (T3/HDF) for 14 passages. A feeder-free culture on Matrigel in hES medium conditioned by these autogeneic feeder cells (T3HDF) was established to maintain the undifferentiated growth of hES-T3 cells (T3/CMHDF) for 8 passages. Results: The gene expression profiles of mRNAs, microRNAs and proteins between the undifferentiated T3/HDF and T3/CMHDF cells were shown to be very similar, and their expression profiles were also found to be similar to those of T3/MEF and T3/CMMEF cells grown on MEF feeder and feeder-free Matrigel in MEF-conditioned medium, respectively. The abundantly expressed genes of T3/HDF, T3/CMHDF, T3/MEF and T3/CMMEF cells were found to play prominent roles in signaling pathways and GO process networks. The miR-302/367 cluster and miR-371/372/373 cluster were extremely abundantly expressed in undifferentiated T3/HDF and T3/CMHDF as well as T3/MEF andT3/CMMEF cells. The undifferentiated state of T3/HDF and T3/CMHDF cells was evidenced by the very high expression levels of ¡§stemness¡¨ genes and low expression levels of differentiation markers of ectoderm, mesoderm and endoderm in addition to the strong staining of OCT4 and NANOG. Conclusion: The T3HDF feeder and T3HDF-conditioned medium were able to support the undifferentiated growth of hES cells, and they would be useful for drug development and toxicity testing in addition to the reduced risks of xenogeneic pathogens when used for medical applications such as cell therapies. Keywords: genetic modification
2011-01-12 | GSE19902 | GEO
Project description:Background: The hES-T3 cell line with normal female karyotype was used to differentiate into autogeneic fibroblast-like cells (T3HDF) as feeder to support the undifferentiated growth of hES-T3 cells (T3/HDF) for 14 passages. A feeder-free culture on Matrigel in hES medium conditioned by these autogeneic feeder cells (T3HDF) was established to maintain the undifferentiated growth of hES-T3 cells (T3/CMHDF) for 8 passages. Results: The gene expression profiles of mRNAs, microRNAs and proteins between the undifferentiated T3/HDF and T3/CMHDF cells were shown to be very similar, and their expression profiles were also found to be similar to those of T3/MEF and T3/CMMEF cells grown on MEF feeder and feeder-free Matrigel in MEF-conditioned medium, respectively. The abundantly expressed genes of T3/HDF, T3/CMHDF, T3/MEF and T3/CMMEF cells were found to play prominent roles in signaling pathways and GO process networks. The miR-302/367 cluster and miR-371/372/373 cluster were extremely abundantly expressed in undifferentiated T3/HDF and T3/CMHDF as well as T3/MEF andT3/CMMEF cells. The undifferentiated state of T3/HDF and T3/CMHDF cells was evidenced by the very high expression levels of ¡§stemness¡¨ genes and low expression levels of differentiation markers of ectoderm, mesoderm and endoderm in addition to the strong staining of OCT4 and NANOG. Conclusion: The T3HDF feeder and T3HDF-conditioned medium were able to support the undifferentiated growth of hES cells, and they would be useful for drug development and toxicity testing in addition to the reduced risks of xenogeneic pathogens when used for medical applications such as cell therapies. Keywords: genetic modification In this investigation, both miRNA and mRNA expression profiles from human embryonic stem cells (hES-T3) grown on MEF feeder (T3_MEF), feeder-free Matrigel in MEF-conditioned medium (T3_CMMEF), T3HDF (hES-T3 differentiated fibroblast-like cells) feeder and feeder-free Matrigel in T3HDF-conditioned medium were quantitatively determined. Several target genes of T3BA and T3CM cells-specific miRNAs were identified. ***This submission represents the mRNA expression component of the study only***
2011-01-12 | E-GEOD-19902 | biostudies-arrayexpress