Unknown,Transcriptomics,Genomics,Proteomics

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MiRNA expression of 3 prostate cell types from human normal donors

ABSTRACT: Prostate primary epithelial and stromal cells were isolated from normal donorhuman prostate tissue and patient prostate tumors (>Gleason 8) using in vitroselective media conditions and differential substrate adhesion properties.Tumor cells specific to the prostate epithelial cell component attached tomatrigel while epithelial cells from donors preferentially attached to gelatinsubstrate. The prostate tumor cells did not proliferate over time in culturebut cells of normal epithelial phenotype in comparison to the donor tissueeventually emerged from this cell compartment. The epithelial componentof both donor and tumor specimens produced stem cell colonies thatexpressed OCT4 in growth factor reduced media but subsequently formedembryoid bodies in hanging drop cultures which differentiated into multiplegerm line lineages under the influence of growth factors. To determineif prostate tumor-derived stem cells were capable of forming tumors invivo, we compared the behavior of the donor and tumor stem cells aftertransplantation as tissue recombinants under the renal capsule of SCID micein the presence of rat urogenital mesenchyme (rUGM). Stem cells from bothdonor and tumor sources produced glandular structures that secreted prostatespecific antigen (PSA) while no glandular structures were formed from tissue recombinants of normal epithelial or tumor epithelial cells plus rUGM or fromrUGM alone. Serial transplantation of the stem cell recombinants from tumor specimens yielded subrenal structures reflecting an abundance of glandswith morphological features typical of Prostatic Intraepithelial Neoplasia(PIN). In order to determine whether intrinsic differences existed amongthe donor-derived and tumor-derived stem cells, array based microRNAexpression profiling was performed on all cell types obtained after in vitro isolation. Sample Labeling Key: PC-Prostate Cancer PD-Prostate Donor (Normal Tissue) DNE-Donor Normal Epithelial NS-Donor Normal Stem TNE-Tumor Normal Epithelial TE-Tumor Epithelial (TNE and TE samples with corresponding numbers are matched pairs) TS-Tumor Stem Cell S-Stromal PC 73 TNE was run in all 6 batches as a control for batch effect. The number in parentheses corresponds with the batch the sample was run in.

ORGANISM(S): Homo sapiens

SUBMITTER: John Krill-Burger

PROVIDER: E-GEOD-17323 | biostudies-arrayexpress |

REPOSITORIES: biostudies-arrayexpress

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Project description:Prostate primary epithelial and stromal cells were isolated from normal donorhuman prostate tissue and patient prostate tumors (>Gleason 8) using in vitroselective media conditions and differential substrate adhesion properties.Tumor cells specific to the prostate epithelial cell component attached tomatrigel while epithelial cells from donors preferentially attached to gelatinsubstrate. The prostate tumor cells did not proliferate over time in culturebut cells of normal epithelial phenotype in comparison to the donor tissueeventually emerged from this cell compartment. The epithelial componentof both donor and tumor specimens produced stem cell colonies thatexpressed OCT4 in growth factor reduced media but subsequently formedembryoid bodies in hanging drop cultures which differentiated into multiplegerm line lineages under the influence of growth factors. To determineif prostate tumor-derived stem cells were capable of forming tumors invivo, we compared the behavior of the donor and tumor stem cells aftertransplantation as tissue recombinants under the renal capsule of SCID micein the presence of rat urogenital mesenchyme (rUGM). Stem cells from bothdonor and tumor sources produced glandular structures that secreted prostatespecific antigen (PSA) while no glandular structures were formed from tissuerecombinants of normal epithelial or tumor epithelial cells plus rUGM or fromrUGM alone. Serial transplantation of the stem cell recombinants from tumorspecimens yielded subrenal structures reflecting an abundance of glands with morphological features typical of Prostatic Intraepithelial Neoplasia(PIN). In order to determine whether intrinsic differences existed amongthe donor-derived and tumor-derived stem cells, array based microRNAexpression profiling was performed on all cell types obtained after in vitro isolation. Sample Labeling Key: PC-Prostate Cancer PD-Prostate Donor (Normal Tissue) DNE-Donor Normal Epithelial NS-Donor Normal Stem TNE-Tumor Normal Epithelial TE-Tumor Epithelial (TNE and TE samples with corresponding numbers are matched pairs) TS-Tumor Stem Cell S-Stromal PC 73 TNE was run in all 6 batches as a control for batch effect. The number in parentheses corresponds with the batch the sample was run in.

Project description:Tissue remodeling or regeneration is believed to initiate from organ-specific stem and/or progenitor cells. We report here the establishment of two spontaneously immortalized adult human benign prostate epithelial cell lines, NHPrE1 and BHPrE1. NHPrE1CD133(++)/CD44(++)/OCT4(++)/CK14(+++) was characterized as a putative progenitor cell, and BHPrE1CD133(+)/CD44(+)/OCT4(+)/CK14(++) as a putative intermediate cell. aCGH analysis demonstrated a largely intact diploid karyotype with DNA amplification of PTEN in NHPrE1 or Beta-Catenin in BHPrE1 cells, respectively. A tissue recombination-xenografting model was utilized to compare remodeling of human prostatic tissues in vivo by the NHPrE1 and BHPrE1 cell lines and their parental primary cells. A series of tissue recombinants, made by mixing different ratios of human prostatic epithelial cells and inductive rat urogenital sinus mesenchyme (UGM), were grafted to the renal capsule of SCID mice. Both cell lines were able to regenerate ductal/acinar structures in vivo containing basal and luminal epithelial populations confirmed by the expression of appropriate cytokeratin profiles. Androgen receptor (AR), prostate specific antigen (PSA), and 15-lipoxigenase-2 (15-LOX-2) expression were appropriately expressed in the regenerated epithelial structures. Regeneration of benign human prostatic ductal/acinar architecture could be achieved using as few as 10 NHPrE1 cells while 200,000 BHPrE1 cells were required to achieve prostatic structure. This suggests a greater proportion of progenitor cells in NHPrE1 facilitated regeneration of prostatic morphology and function. These cell lines provide important data on progenitor and intermediate cell phenotypes and represent significant new tools for the elucidation of molecular mechanisms of human prostatic regeneration, pathogenesis and carcinogenesis. Two channel CGH from Agilent platform was used to detect DNA amplification or deletion in genomic samples from NHPrE1 and BHPrE1 cell lines. A common pooled normal DNA refernce was used.

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MiRNA expression of 3 prostate cell types from human normal donors

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