Progenitor Cell Line (hPheo1) Derived from a Human Pheochromocytoma Tumor

Hans K. Ghayee, Vikash J. Bhagwandin, Victor Stastny, Arielle Click, Liang Hao Ding, Dario Mizrachi, Ying S. Zou, Raj Chari, Wan L. Lam, Robert M. Bachoo, Alice L. Smith, Michael D. Story, Stan Sidhu, Bruce G. Robinson, Fiemu E. Nwariaku, Adi F. Gazdar, Richard J. Auchus, Jerry W. Shay

Research output: Contribution to journalArticle

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Abstract

Background:Pheochromocytomas are rare tumors generally arising in the medullary region of the adrenal gland. These tumors release excessive epinephrine and norepinephrine resulting in hypertension and cardiovascular crises for which surgery is the only definitive treatment. Molecular mechanisms that control tumor development and hormone production are poorly understood, and progress has been hampered by the lack of human cellular model systems. To study pheochromocytomas, we developed a stable progenitor pheochromocytoma cell line derived from a primary human tumor.Methods:After IRB approval and written informed consent, human pheochromocytoma tissue was excised, minced, dispersed enzymatically, and cultured in vitro. Primary pheochromocytoma cells were infected with a lentivirus vector carrying the catalytic subunit of human telomerase reverse transcriptase (hTERT). The hTERT immortalized cells (hPheo1) have been passaged >300 population doublings. The resulting cell line was characterized morphologically, biochemically and for expression of neuroendocrine properties. The expression of marker enzymes and proteins was assessed by immunofluorescence staining and immunoblotting. Telomerase activity was determined by using the telomeric repeat amplification protocol (TRAP) assay.Results:We have established a human pheochromocytoma precursor cell line that expresses the neuroendocrine marker, chromogranin A, when differentiated in the presence of bone morphogenic protein 4 (BMP4), nerve growth factor (NGF), and dexamethasone. Phenylethanolamine N-methyltransferase (PNMT) expression is also detected with this differentiation regimen. CD-56 (also known as NCAM, neural cell adhesion molecule) is expressed in these cells, but CD31 (also known as PECAM-1, a marker of endothelial cells) is negative.Conclusions:We have maintained hTERT-immortalized progenitor cells derived from a pheochromocytoma (hPheo1) in culture for over 300 population doublings. This progenitor human cell line is normal diploid except for a deletion in the p16 region and has inducible neuroendocrine biomarkers. These cells should be a valuable reagent for studying mechanisms of tumor development and for testing novel therapeutic approaches.

Original languageEnglish (US)
Article numbere65624
JournalPLoS One
Volume8
Issue number6
DOIs
StatePublished - Jun 13 2013

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Pheochromocytoma
stem cells
Tumors
Stem Cells
Cells
cell lines
telomerase
Cell Line
neoplasms
Neural Cell Adhesion Molecules
RNA-directed DNA polymerase
PC12 Cells
Neoplasms
Phenylethanolamine N-Methyltransferase
CD31 Antigens
Chromogranin A
Lentivirus
phenylethanolamine N-methyltransferase
Research Ethics Committees
Telomerase

ASJC Scopus subject areas

  • Agricultural and Biological Sciences(all)
  • Biochemistry, Genetics and Molecular Biology(all)
  • Medicine(all)

Cite this

Progenitor Cell Line (hPheo1) Derived from a Human Pheochromocytoma Tumor. / Ghayee, Hans K.; Bhagwandin, Vikash J.; Stastny, Victor; Click, Arielle; Ding, Liang Hao; Mizrachi, Dario; Zou, Ying S.; Chari, Raj; Lam, Wan L.; Bachoo, Robert M.; Smith, Alice L.; Story, Michael D.; Sidhu, Stan; Robinson, Bruce G.; Nwariaku, Fiemu E.; Gazdar, Adi F.; Auchus, Richard J.; Shay, Jerry W.

In: PLoS One, Vol. 8, No. 6, e65624, 13.06.2013.

Research output: Contribution to journalArticle

Ghayee, HK, Bhagwandin, VJ, Stastny, V, Click, A, Ding, LH, Mizrachi, D, Zou, YS, Chari, R, Lam, WL, Bachoo, RM, Smith, AL, Story, MD, Sidhu, S, Robinson, BG, Nwariaku, FE, Gazdar, AF, Auchus, RJ & Shay, JW 2013, 'Progenitor Cell Line (hPheo1) Derived from a Human Pheochromocytoma Tumor', PLoS One, vol. 8, no. 6, e65624. https://doi.org/10.1371/journal.pone.0065624
Ghayee HK, Bhagwandin VJ, Stastny V, Click A, Ding LH, Mizrachi D et al. Progenitor Cell Line (hPheo1) Derived from a Human Pheochromocytoma Tumor. PLoS One. 2013 Jun 13;8(6). e65624. https://doi.org/10.1371/journal.pone.0065624
Ghayee, Hans K. ; Bhagwandin, Vikash J. ; Stastny, Victor ; Click, Arielle ; Ding, Liang Hao ; Mizrachi, Dario ; Zou, Ying S. ; Chari, Raj ; Lam, Wan L. ; Bachoo, Robert M. ; Smith, Alice L. ; Story, Michael D. ; Sidhu, Stan ; Robinson, Bruce G. ; Nwariaku, Fiemu E. ; Gazdar, Adi F. ; Auchus, Richard J. ; Shay, Jerry W. / Progenitor Cell Line (hPheo1) Derived from a Human Pheochromocytoma Tumor. In: PLoS One. 2013 ; Vol. 8, No. 6.
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abstract = "Background:Pheochromocytomas are rare tumors generally arising in the medullary region of the adrenal gland. These tumors release excessive epinephrine and norepinephrine resulting in hypertension and cardiovascular crises for which surgery is the only definitive treatment. Molecular mechanisms that control tumor development and hormone production are poorly understood, and progress has been hampered by the lack of human cellular model systems. To study pheochromocytomas, we developed a stable progenitor pheochromocytoma cell line derived from a primary human tumor.Methods:After IRB approval and written informed consent, human pheochromocytoma tissue was excised, minced, dispersed enzymatically, and cultured in vitro. Primary pheochromocytoma cells were infected with a lentivirus vector carrying the catalytic subunit of human telomerase reverse transcriptase (hTERT). The hTERT immortalized cells (hPheo1) have been passaged >300 population doublings. The resulting cell line was characterized morphologically, biochemically and for expression of neuroendocrine properties. The expression of marker enzymes and proteins was assessed by immunofluorescence staining and immunoblotting. Telomerase activity was determined by using the telomeric repeat amplification protocol (TRAP) assay.Results:We have established a human pheochromocytoma precursor cell line that expresses the neuroendocrine marker, chromogranin A, when differentiated in the presence of bone morphogenic protein 4 (BMP4), nerve growth factor (NGF), and dexamethasone. Phenylethanolamine N-methyltransferase (PNMT) expression is also detected with this differentiation regimen. CD-56 (also known as NCAM, neural cell adhesion molecule) is expressed in these cells, but CD31 (also known as PECAM-1, a marker of endothelial cells) is negative.Conclusions:We have maintained hTERT-immortalized progenitor cells derived from a pheochromocytoma (hPheo1) in culture for over 300 population doublings. This progenitor human cell line is normal diploid except for a deletion in the p16 region and has inducible neuroendocrine biomarkers. These cells should be a valuable reagent for studying mechanisms of tumor development and for testing novel therapeutic approaches.",
author = "Ghayee, {Hans K.} and Bhagwandin, {Vikash J.} and Victor Stastny and Arielle Click and Ding, {Liang Hao} and Dario Mizrachi and Zou, {Ying S.} and Raj Chari and Lam, {Wan L.} and Bachoo, {Robert M.} and Smith, {Alice L.} and Story, {Michael D.} and Stan Sidhu and Robinson, {Bruce G.} and Nwariaku, {Fiemu E.} and Gazdar, {Adi F.} and Auchus, {Richard J.} and Shay, {Jerry W.}",
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T1 - Progenitor Cell Line (hPheo1) Derived from a Human Pheochromocytoma Tumor

AU - Ghayee, Hans K.

AU - Bhagwandin, Vikash J.

AU - Stastny, Victor

AU - Click, Arielle

AU - Ding, Liang Hao

AU - Mizrachi, Dario

AU - Zou, Ying S.

AU - Chari, Raj

AU - Lam, Wan L.

AU - Bachoo, Robert M.

AU - Smith, Alice L.

AU - Story, Michael D.

AU - Sidhu, Stan

AU - Robinson, Bruce G.

AU - Nwariaku, Fiemu E.

AU - Gazdar, Adi F.

AU - Auchus, Richard J.

AU - Shay, Jerry W.

PY - 2013/6/13

Y1 - 2013/6/13

N2 - Background:Pheochromocytomas are rare tumors generally arising in the medullary region of the adrenal gland. These tumors release excessive epinephrine and norepinephrine resulting in hypertension and cardiovascular crises for which surgery is the only definitive treatment. Molecular mechanisms that control tumor development and hormone production are poorly understood, and progress has been hampered by the lack of human cellular model systems. To study pheochromocytomas, we developed a stable progenitor pheochromocytoma cell line derived from a primary human tumor.Methods:After IRB approval and written informed consent, human pheochromocytoma tissue was excised, minced, dispersed enzymatically, and cultured in vitro. Primary pheochromocytoma cells were infected with a lentivirus vector carrying the catalytic subunit of human telomerase reverse transcriptase (hTERT). The hTERT immortalized cells (hPheo1) have been passaged >300 population doublings. The resulting cell line was characterized morphologically, biochemically and for expression of neuroendocrine properties. The expression of marker enzymes and proteins was assessed by immunofluorescence staining and immunoblotting. Telomerase activity was determined by using the telomeric repeat amplification protocol (TRAP) assay.Results:We have established a human pheochromocytoma precursor cell line that expresses the neuroendocrine marker, chromogranin A, when differentiated in the presence of bone morphogenic protein 4 (BMP4), nerve growth factor (NGF), and dexamethasone. Phenylethanolamine N-methyltransferase (PNMT) expression is also detected with this differentiation regimen. CD-56 (also known as NCAM, neural cell adhesion molecule) is expressed in these cells, but CD31 (also known as PECAM-1, a marker of endothelial cells) is negative.Conclusions:We have maintained hTERT-immortalized progenitor cells derived from a pheochromocytoma (hPheo1) in culture for over 300 population doublings. This progenitor human cell line is normal diploid except for a deletion in the p16 region and has inducible neuroendocrine biomarkers. These cells should be a valuable reagent for studying mechanisms of tumor development and for testing novel therapeutic approaches.

AB - Background:Pheochromocytomas are rare tumors generally arising in the medullary region of the adrenal gland. These tumors release excessive epinephrine and norepinephrine resulting in hypertension and cardiovascular crises for which surgery is the only definitive treatment. Molecular mechanisms that control tumor development and hormone production are poorly understood, and progress has been hampered by the lack of human cellular model systems. To study pheochromocytomas, we developed a stable progenitor pheochromocytoma cell line derived from a primary human tumor.Methods:After IRB approval and written informed consent, human pheochromocytoma tissue was excised, minced, dispersed enzymatically, and cultured in vitro. Primary pheochromocytoma cells were infected with a lentivirus vector carrying the catalytic subunit of human telomerase reverse transcriptase (hTERT). The hTERT immortalized cells (hPheo1) have been passaged >300 population doublings. The resulting cell line was characterized morphologically, biochemically and for expression of neuroendocrine properties. The expression of marker enzymes and proteins was assessed by immunofluorescence staining and immunoblotting. Telomerase activity was determined by using the telomeric repeat amplification protocol (TRAP) assay.Results:We have established a human pheochromocytoma precursor cell line that expresses the neuroendocrine marker, chromogranin A, when differentiated in the presence of bone morphogenic protein 4 (BMP4), nerve growth factor (NGF), and dexamethasone. Phenylethanolamine N-methyltransferase (PNMT) expression is also detected with this differentiation regimen. CD-56 (also known as NCAM, neural cell adhesion molecule) is expressed in these cells, but CD31 (also known as PECAM-1, a marker of endothelial cells) is negative.Conclusions:We have maintained hTERT-immortalized progenitor cells derived from a pheochromocytoma (hPheo1) in culture for over 300 population doublings. This progenitor human cell line is normal diploid except for a deletion in the p16 region and has inducible neuroendocrine biomarkers. These cells should be a valuable reagent for studying mechanisms of tumor development and for testing novel therapeutic approaches.

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