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Functional remodeling of benign human prostatic tissues in vivo by spontaneously immortalized progenitor and intermediate cells.

Jiang M, Strand DW, Fernandez S, He Y, Yi Y, Birbach A, Qiu Q, Schmid J, Tang DG, Hayward SW - Stem Cells (2010)

Bottom Line: Embedded three-dimensional culture of NHPrE1 showed greater branching than BHPrE1.Regeneration of benign prostatic glandular structures could be achieved using as few as 10 NHPrE1 cells, whereas 200,000 BHPrE1 cells were required to achieve prostatic architecture.This suggests a greater proportion of progenitor/stem cells in NHPrE1 than in BHPrE1.

View Article: PubMed Central - PubMed

Affiliation: Department of Urological Surgery, Vanderbilt University Medical Center and Vanderbilt-Ingram Cancer Center, Nashville, Tennessee 37232-2765, USA. ming.jiang.1@vanderbilt.edu

ABSTRACT
Tissue remodeling or regeneration is believed to initiate from multipotent stem and progenitor cells. We report here the establishment of two spontaneously immortalized adult non-tumorigenic human prostate epithelial cell lines, NHPrE1 and BHPrE1. NHPrE1 (CD133(high)/CD44(high)/OCT4(high)/PTEN(high)) was characterized as a putative progenitor cell, and BHPrE1 (p63(high)/p53(high)/p21(WAF1)(high)/RB(high)) was characterized as a putative epithelial intermediate cell. Genomic analysis demonstrated an abnormal karyotype with genomic rearrangements including PTEN amplification in NHPrE1 and CTNNB1 (beta-catenin) amplification in BHPrE1 cells. Embedded three-dimensional culture of NHPrE1 showed greater branching than BHPrE1. A tissue recombination-xenografting model was utilized to compare remodeling of human prostatic tissues in vivo. A series of tissue recombinants, made by mixing different ratios of human prostatic epithelial cells and inductive rat urogenital sinus mesenchyme, were grafted to the renal capsule of severe combined immunodeficient mice. Both cell lines were able to regenerate benign secretory ductal-acinar architecture in vivo, containing intact basal and luminal epithelial layers confirmed by the expression of appropriate CK profiles. Prostate-specific antigen, 15-lipoxygenase-2, androgen receptor, and NKX3.1 proteins were appropriately expressed in the regenerated epithelia. Regeneration of benign prostatic glandular structures could be achieved using as few as 10 NHPrE1 cells, whereas 200,000 BHPrE1 cells were required to achieve prostatic architecture. This suggests a greater proportion of progenitor/stem cells in NHPrE1 than in BHPrE1. 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.

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Array–comparative genomic hybridization (array-CGH) and differential gene locus mapping (DIGMAP) analysis of NHPrE0 and BHPrE0 primary cells and derivative NHPrE1 and BHPrE1 cell lines. A: Array-CGH of NHPrE0 and BHPrE0 lines reveal a genome with no prominent unbalanced events; karyotypes resemble “normal.” In contrast, the derivative cell lines (BHPrE1 and NHPrE1) show more extensive genomic rearrangements. B: Converted DIGMAP data showing alterations in PTEN (amplified in NHPrE1), CTNNB1 (β-catenin amplified in BHPrE1), TERT, CDKN2A (p16), and TP53, which facilitate spontaneous immortalization in both lines. All array-CGH data have been deposited at the GEO database (accession number GSE18122). Abbreviations: BHPrE0, primary cultured benign human prostate epithelial cells isolated from a prostate surgical sample; BHPrE1, a spontaneously immortalized benign human prostate epithelial cell line from BHPrE0; NHPrE0 (NHP8), primary cultured “normal” human prostate epithelial cells isolated from a donated, 41-year-old, healthy, male prostate; NHPrE1: a spontaneously immortalized “normal” human prostate epithelial cell line, that was established from parent NHPrE0 cells.
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fig04: Array–comparative genomic hybridization (array-CGH) and differential gene locus mapping (DIGMAP) analysis of NHPrE0 and BHPrE0 primary cells and derivative NHPrE1 and BHPrE1 cell lines. A: Array-CGH of NHPrE0 and BHPrE0 lines reveal a genome with no prominent unbalanced events; karyotypes resemble “normal.” In contrast, the derivative cell lines (BHPrE1 and NHPrE1) show more extensive genomic rearrangements. B: Converted DIGMAP data showing alterations in PTEN (amplified in NHPrE1), CTNNB1 (β-catenin amplified in BHPrE1), TERT, CDKN2A (p16), and TP53, which facilitate spontaneous immortalization in both lines. All array-CGH data have been deposited at the GEO database (accession number GSE18122). Abbreviations: BHPrE0, primary cultured benign human prostate epithelial cells isolated from a prostate surgical sample; BHPrE1, a spontaneously immortalized benign human prostate epithelial cell line from BHPrE0; NHPrE0 (NHP8), primary cultured “normal” human prostate epithelial cells isolated from a donated, 41-year-old, healthy, male prostate; NHPrE1: a spontaneously immortalized “normal” human prostate epithelial cell line, that was established from parent NHPrE0 cells.

Mentions: With long-term continuous culture in the serum-supplemented HPrE-conditional medium, NHPrE1 cells showed a near diploid karyotype with a 3n sideline. BHPrE1 cells were near 3n, 65–72 chromosomes with significant chromosomal rearrangements. Array-CGH was used to detect DNA amplification or deletion in genomic samples from NHPrE1 and BHPrE1 cell lines. Global views of the array-CGH results for NHPrE1 and BHPrE1 cell lines, their parental NHPrE0 and BHPrE0 cells were plotted (Fig. 4A). There was an obvious hemizygous deletion of TP53, but not WAF1 (p21), Cyclin D1, or RB, in both NHPrE1 and BHPrE1 cells, compared to the array-CGH results for the parental NHPrE0 and BHPrE0 cells (which were apparently genetically intact with minimal chromosomal changes). The copy number changes for chromosome regions were examined using DIGMAP analysis to compare the four cell types (NHPrE0, NHPrE1, BPHPrE0, and BHPrE1) as shown in Figure 4B. The main differential regions between NHPrE1 and BHPrE1 are 17p13.1, 10q23.3, 3p21, 5p15.33, and 9p21. Interestingly, PTEN was amplified in NHPrE1 cells and CTNNB1 (β-catenin) was amplified in BHPrE1 cells (Fig. 4B). These data support the concept that PTEN may be involved in some way in maintaining progenitor cell status [29]. TERT, CDKN2A (p16), and TP53 status in both cell lines (Fig. 4B) also supported the observed phenotypes during cellular immortalization [19]. An extended DIGMAP analysis of additional progenitor and differentiation markers are listed in supporting information.


Functional remodeling of benign human prostatic tissues in vivo by spontaneously immortalized progenitor and intermediate cells.

Jiang M, Strand DW, Fernandez S, He Y, Yi Y, Birbach A, Qiu Q, Schmid J, Tang DG, Hayward SW - Stem Cells (2010)

Array–comparative genomic hybridization (array-CGH) and differential gene locus mapping (DIGMAP) analysis of NHPrE0 and BHPrE0 primary cells and derivative NHPrE1 and BHPrE1 cell lines. A: Array-CGH of NHPrE0 and BHPrE0 lines reveal a genome with no prominent unbalanced events; karyotypes resemble “normal.” In contrast, the derivative cell lines (BHPrE1 and NHPrE1) show more extensive genomic rearrangements. B: Converted DIGMAP data showing alterations in PTEN (amplified in NHPrE1), CTNNB1 (β-catenin amplified in BHPrE1), TERT, CDKN2A (p16), and TP53, which facilitate spontaneous immortalization in both lines. All array-CGH data have been deposited at the GEO database (accession number GSE18122). Abbreviations: BHPrE0, primary cultured benign human prostate epithelial cells isolated from a prostate surgical sample; BHPrE1, a spontaneously immortalized benign human prostate epithelial cell line from BHPrE0; NHPrE0 (NHP8), primary cultured “normal” human prostate epithelial cells isolated from a donated, 41-year-old, healthy, male prostate; NHPrE1: a spontaneously immortalized “normal” human prostate epithelial cell line, that was established from parent NHPrE0 cells.
© Copyright Policy - open-access
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC2962907&req=5

fig04: Array–comparative genomic hybridization (array-CGH) and differential gene locus mapping (DIGMAP) analysis of NHPrE0 and BHPrE0 primary cells and derivative NHPrE1 and BHPrE1 cell lines. A: Array-CGH of NHPrE0 and BHPrE0 lines reveal a genome with no prominent unbalanced events; karyotypes resemble “normal.” In contrast, the derivative cell lines (BHPrE1 and NHPrE1) show more extensive genomic rearrangements. B: Converted DIGMAP data showing alterations in PTEN (amplified in NHPrE1), CTNNB1 (β-catenin amplified in BHPrE1), TERT, CDKN2A (p16), and TP53, which facilitate spontaneous immortalization in both lines. All array-CGH data have been deposited at the GEO database (accession number GSE18122). Abbreviations: BHPrE0, primary cultured benign human prostate epithelial cells isolated from a prostate surgical sample; BHPrE1, a spontaneously immortalized benign human prostate epithelial cell line from BHPrE0; NHPrE0 (NHP8), primary cultured “normal” human prostate epithelial cells isolated from a donated, 41-year-old, healthy, male prostate; NHPrE1: a spontaneously immortalized “normal” human prostate epithelial cell line, that was established from parent NHPrE0 cells.
Mentions: With long-term continuous culture in the serum-supplemented HPrE-conditional medium, NHPrE1 cells showed a near diploid karyotype with a 3n sideline. BHPrE1 cells were near 3n, 65–72 chromosomes with significant chromosomal rearrangements. Array-CGH was used to detect DNA amplification or deletion in genomic samples from NHPrE1 and BHPrE1 cell lines. Global views of the array-CGH results for NHPrE1 and BHPrE1 cell lines, their parental NHPrE0 and BHPrE0 cells were plotted (Fig. 4A). There was an obvious hemizygous deletion of TP53, but not WAF1 (p21), Cyclin D1, or RB, in both NHPrE1 and BHPrE1 cells, compared to the array-CGH results for the parental NHPrE0 and BHPrE0 cells (which were apparently genetically intact with minimal chromosomal changes). The copy number changes for chromosome regions were examined using DIGMAP analysis to compare the four cell types (NHPrE0, NHPrE1, BPHPrE0, and BHPrE1) as shown in Figure 4B. The main differential regions between NHPrE1 and BHPrE1 are 17p13.1, 10q23.3, 3p21, 5p15.33, and 9p21. Interestingly, PTEN was amplified in NHPrE1 cells and CTNNB1 (β-catenin) was amplified in BHPrE1 cells (Fig. 4B). These data support the concept that PTEN may be involved in some way in maintaining progenitor cell status [29]. TERT, CDKN2A (p16), and TP53 status in both cell lines (Fig. 4B) also supported the observed phenotypes during cellular immortalization [19]. An extended DIGMAP analysis of additional progenitor and differentiation markers are listed in supporting information.

Bottom Line: Embedded three-dimensional culture of NHPrE1 showed greater branching than BHPrE1.Regeneration of benign prostatic glandular structures could be achieved using as few as 10 NHPrE1 cells, whereas 200,000 BHPrE1 cells were required to achieve prostatic architecture.This suggests a greater proportion of progenitor/stem cells in NHPrE1 than in BHPrE1.

View Article: PubMed Central - PubMed

Affiliation: Department of Urological Surgery, Vanderbilt University Medical Center and Vanderbilt-Ingram Cancer Center, Nashville, Tennessee 37232-2765, USA. ming.jiang.1@vanderbilt.edu

ABSTRACT
Tissue remodeling or regeneration is believed to initiate from multipotent stem and progenitor cells. We report here the establishment of two spontaneously immortalized adult non-tumorigenic human prostate epithelial cell lines, NHPrE1 and BHPrE1. NHPrE1 (CD133(high)/CD44(high)/OCT4(high)/PTEN(high)) was characterized as a putative progenitor cell, and BHPrE1 (p63(high)/p53(high)/p21(WAF1)(high)/RB(high)) was characterized as a putative epithelial intermediate cell. Genomic analysis demonstrated an abnormal karyotype with genomic rearrangements including PTEN amplification in NHPrE1 and CTNNB1 (beta-catenin) amplification in BHPrE1 cells. Embedded three-dimensional culture of NHPrE1 showed greater branching than BHPrE1. A tissue recombination-xenografting model was utilized to compare remodeling of human prostatic tissues in vivo. A series of tissue recombinants, made by mixing different ratios of human prostatic epithelial cells and inductive rat urogenital sinus mesenchyme, were grafted to the renal capsule of severe combined immunodeficient mice. Both cell lines were able to regenerate benign secretory ductal-acinar architecture in vivo, containing intact basal and luminal epithelial layers confirmed by the expression of appropriate CK profiles. Prostate-specific antigen, 15-lipoxygenase-2, androgen receptor, and NKX3.1 proteins were appropriately expressed in the regenerated epithelia. Regeneration of benign prostatic glandular structures could be achieved using as few as 10 NHPrE1 cells, whereas 200,000 BHPrE1 cells were required to achieve prostatic architecture. This suggests a greater proportion of progenitor/stem cells in NHPrE1 than in BHPrE1. 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.

Show MeSH
Related in: MedlinePlus