Pitfalls in Detection of Contaminating Neuroblastoma Cells by Tyrosine Hydroxylase RT-PCR Due to Catecholamine-producing Hematopoietic Cells
Background: RT-PCR analysis of compounds of catecholamine metabolism (in particular tyrosine hydroxylase, TH) is widely used for the detection of contaminating neuroblastoma cells in hematopoietic stem cell preparations. Due to reports in the literature showing that hematopoietic cells are also able...
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| Veröffentlicht in: | Anticancer research 2006-05, Vol.26 (3A), p.2075-2080 |
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| creator | KUCI, Zyrafete SEITZ, Gabriele KUCI, Selim KREYENBERG, Hermann SCHUMM, Michael LANG, Peter NIETHAMMER, Dietrich HANDGRETINGER, Rupert BRUCHELT, Gernot |
| description | Background: RT-PCR analysis of compounds of catecholamine metabolism (in particular tyrosine hydroxylase, TH) is widely used
for the detection of contaminating neuroblastoma cells in hematopoietic stem cell preparations. Due to reports in the literature
showing that hematopoietic cells are also able to produce catecholamines, we investigated whether TH-RT-PCR is really suitable
for this purpose. Materials and Methods: Besides neuroblastoma cells, mononuclear blood cells, apheresis preparations and
hematopoietic stem cells were used for single and nested RT-PCR. In addition to TH, the expressions of dopamine-β-hydroxylase
and noradrenaline transporter were analyzed. Results: Using single RT-PCR, a clear discrimination between neuroblastoma and
hematopoietic cells was possible. However, by using nested RT-PCR, the âneuroblastoma markersâ were also detected in a significant
percentage of non-mobilized mononuclear blood cells, in mononuclear blood cells of healthy donors mobilized with G-CSF, and
in highly purified CD34 + and CD133 + stem cells from healthy mobilized donors. Conclusion: Our results raise the question of whether the RT-PCR analysis of compounds
of catecholamine metabolism is suitable and selective enough to detect the contamination of hematopoietic stem cells by a
low number of neuroblastoma cells. |
| format | Article |
| fullrecord | <record><control><sourceid>proquest_pubme</sourceid><recordid>TN_cdi_proquest_miscellaneous_68613429</recordid><sourceformat>XML</sourceformat><sourcesystem>PC</sourcesystem><sourcerecordid>68613429</sourcerecordid><originalsourceid>FETCH-LOGICAL-h270t-ab955fbe5cfb3735f65e2b91845b9e022ecb423823b9956454b1d761c06243783</originalsourceid><addsrcrecordid>eNpFkctOwzAQRSMEouXxC8gb2EVy7NiOl1V4FAkBQmUd2e6EGiVxsR1B_oGPxhVFrGZz5szVnYNsXghZ5IJRfJjNMWE4FxizWXYSwjvGnMuKHmezgldEFKWYZ9_PNraq6wKyA7qGCCZaNyDXotoNUfV2UNEOb-gRRu90p0J0vUI17Db0hFaTd8EOgJbT2ruvKQGAXlb5c_2CrkdA0aFaJenGdTsX5Fvv1qPZGZfQq-i2zkK05td4lh2lLAHO9_M0e729WdXL_OHp7r5ePOQbInDMlZaMtRqYaTUVlLWcAdGyqEqmJWBCwOiS0IpQLSXjJSt1sRa8MJiTkoqKnmZXv96U5mOEEJveBpMSqAHcGBpe8YKWRCbwYg-Ouod1s_W2V35q_vpLwOUeUMGorvVqMDb8c0JiTGT5f3Fj3zaf1kMT-tR60tJGecIbumgITm_7AS0-iHQ</addsrcrecordid><sourcetype>Aggregation Database</sourcetype><iscdi>true</iscdi><recordtype>article</recordtype><pqid>68613429</pqid></control><display><type>article</type><title>Pitfalls in Detection of Contaminating Neuroblastoma Cells by Tyrosine Hydroxylase RT-PCR Due to Catecholamine-producing Hematopoietic Cells</title><source>MEDLINE</source><source>EZB-FREE-00999 freely available EZB journals</source><creator>KUCI, Zyrafete ; SEITZ, Gabriele ; KUCI, Selim ; KREYENBERG, Hermann ; SCHUMM, Michael ; LANG, Peter ; NIETHAMMER, Dietrich ; HANDGRETINGER, Rupert ; BRUCHELT, Gernot</creator><creatorcontrib>KUCI, Zyrafete ; SEITZ, Gabriele ; KUCI, Selim ; KREYENBERG, Hermann ; SCHUMM, Michael ; LANG, Peter ; NIETHAMMER, Dietrich ; HANDGRETINGER, Rupert ; BRUCHELT, Gernot</creatorcontrib><description>Background: RT-PCR analysis of compounds of catecholamine metabolism (in particular tyrosine hydroxylase, TH) is widely used
for the detection of contaminating neuroblastoma cells in hematopoietic stem cell preparations. Due to reports in the literature
showing that hematopoietic cells are also able to produce catecholamines, we investigated whether TH-RT-PCR is really suitable
for this purpose. Materials and Methods: Besides neuroblastoma cells, mononuclear blood cells, apheresis preparations and
hematopoietic stem cells were used for single and nested RT-PCR. In addition to TH, the expressions of dopamine-β-hydroxylase
and noradrenaline transporter were analyzed. Results: Using single RT-PCR, a clear discrimination between neuroblastoma and
hematopoietic cells was possible. However, by using nested RT-PCR, the âneuroblastoma markersâ were also detected in a significant
percentage of non-mobilized mononuclear blood cells, in mononuclear blood cells of healthy donors mobilized with G-CSF, and
in highly purified CD34 + and CD133 + stem cells from healthy mobilized donors. Conclusion: Our results raise the question of whether the RT-PCR analysis of compounds
of catecholamine metabolism is suitable and selective enough to detect the contamination of hematopoietic stem cells by a
low number of neuroblastoma cells.</description><identifier>ISSN: 0250-7005</identifier><identifier>EISSN: 1791-7530</identifier><identifier>PMID: 16827147</identifier><language>eng</language><publisher>Attiki: International Institute of Anticancer Research</publisher><subject>Biological and medical sciences ; Catecholamines - biosynthesis ; Cell Line, Tumor ; Dopamine beta-Hydroxylase - analysis ; Dopamine beta-Hydroxylase - biosynthesis ; Dopamine beta-Hydroxylase - genetics ; Gene Expression Profiling ; Hematopoietic Stem Cells - cytology ; Hematopoietic Stem Cells - enzymology ; Hematopoietic Stem Cells - metabolism ; Humans ; Leukocytes, Mononuclear - cytology ; Leukocytes, Mononuclear - enzymology ; Leukocytes, Mononuclear - metabolism ; Medical sciences ; Neuroblastoma - enzymology ; Neuroblastoma - genetics ; Neuroblastoma - pathology ; Neurology ; Reverse Transcriptase Polymerase Chain Reaction ; Sensitivity and Specificity ; Tumors ; Tumors of the nervous system. Phacomatoses ; Tyrosine 3-Monooxygenase - analysis ; Tyrosine 3-Monooxygenase - biosynthesis ; Tyrosine 3-Monooxygenase - genetics</subject><ispartof>Anticancer research, 2006-05, Vol.26 (3A), p.2075-2080</ispartof><rights>2006 INIST-CNRS</rights><lds50>peer_reviewed</lds50><woscitedreferencessubscribed>false</woscitedreferencessubscribed></display><links><openurl>$$Topenurl_article</openurl><openurlfulltext>$$Topenurlfull_article</openurlfulltext><thumbnail>$$Tsyndetics_thumb_exl</thumbnail><link.rule.ids>314,780,784</link.rule.ids><backlink>$$Uhttp://pascal-francis.inist.fr/vibad/index.php?action=getRecordDetail&idt=17900294$$DView record in Pascal Francis$$Hfree_for_read</backlink><backlink>$$Uhttps://www.ncbi.nlm.nih.gov/pubmed/16827147$$D View this record in MEDLINE/PubMed$$Hfree_for_read</backlink></links><search><creatorcontrib>KUCI, Zyrafete</creatorcontrib><creatorcontrib>SEITZ, Gabriele</creatorcontrib><creatorcontrib>KUCI, Selim</creatorcontrib><creatorcontrib>KREYENBERG, Hermann</creatorcontrib><creatorcontrib>SCHUMM, Michael</creatorcontrib><creatorcontrib>LANG, Peter</creatorcontrib><creatorcontrib>NIETHAMMER, Dietrich</creatorcontrib><creatorcontrib>HANDGRETINGER, Rupert</creatorcontrib><creatorcontrib>BRUCHELT, Gernot</creatorcontrib><title>Pitfalls in Detection of Contaminating Neuroblastoma Cells by Tyrosine Hydroxylase RT-PCR Due to Catecholamine-producing Hematopoietic Cells</title><title>Anticancer research</title><addtitle>Anticancer Res</addtitle><description>Background: RT-PCR analysis of compounds of catecholamine metabolism (in particular tyrosine hydroxylase, TH) is widely used
for the detection of contaminating neuroblastoma cells in hematopoietic stem cell preparations. Due to reports in the literature
showing that hematopoietic cells are also able to produce catecholamines, we investigated whether TH-RT-PCR is really suitable
for this purpose. Materials and Methods: Besides neuroblastoma cells, mononuclear blood cells, apheresis preparations and
hematopoietic stem cells were used for single and nested RT-PCR. In addition to TH, the expressions of dopamine-β-hydroxylase
and noradrenaline transporter were analyzed. Results: Using single RT-PCR, a clear discrimination between neuroblastoma and
hematopoietic cells was possible. However, by using nested RT-PCR, the âneuroblastoma markersâ were also detected in a significant
percentage of non-mobilized mononuclear blood cells, in mononuclear blood cells of healthy donors mobilized with G-CSF, and
in highly purified CD34 + and CD133 + stem cells from healthy mobilized donors. Conclusion: Our results raise the question of whether the RT-PCR analysis of compounds
of catecholamine metabolism is suitable and selective enough to detect the contamination of hematopoietic stem cells by a
low number of neuroblastoma cells.</description><subject>Biological and medical sciences</subject><subject>Catecholamines - biosynthesis</subject><subject>Cell Line, Tumor</subject><subject>Dopamine beta-Hydroxylase - analysis</subject><subject>Dopamine beta-Hydroxylase - biosynthesis</subject><subject>Dopamine beta-Hydroxylase - genetics</subject><subject>Gene Expression Profiling</subject><subject>Hematopoietic Stem Cells - cytology</subject><subject>Hematopoietic Stem Cells - enzymology</subject><subject>Hematopoietic Stem Cells - metabolism</subject><subject>Humans</subject><subject>Leukocytes, Mononuclear - cytology</subject><subject>Leukocytes, Mononuclear - enzymology</subject><subject>Leukocytes, Mononuclear - metabolism</subject><subject>Medical sciences</subject><subject>Neuroblastoma - enzymology</subject><subject>Neuroblastoma - genetics</subject><subject>Neuroblastoma - pathology</subject><subject>Neurology</subject><subject>Reverse Transcriptase Polymerase Chain Reaction</subject><subject>Sensitivity and Specificity</subject><subject>Tumors</subject><subject>Tumors of the nervous system. Phacomatoses</subject><subject>Tyrosine 3-Monooxygenase - analysis</subject><subject>Tyrosine 3-Monooxygenase - biosynthesis</subject><subject>Tyrosine 3-Monooxygenase - genetics</subject><issn>0250-7005</issn><issn>1791-7530</issn><fulltext>true</fulltext><rsrctype>article</rsrctype><creationdate>2006</creationdate><recordtype>article</recordtype><sourceid>EIF</sourceid><recordid>eNpFkctOwzAQRSMEouXxC8gb2EVy7NiOl1V4FAkBQmUd2e6EGiVxsR1B_oGPxhVFrGZz5szVnYNsXghZ5IJRfJjNMWE4FxizWXYSwjvGnMuKHmezgldEFKWYZ9_PNraq6wKyA7qGCCZaNyDXotoNUfV2UNEOb-gRRu90p0J0vUI17Db0hFaTd8EOgJbT2ruvKQGAXlb5c_2CrkdA0aFaJenGdTsX5Fvv1qPZGZfQq-i2zkK05td4lh2lLAHO9_M0e729WdXL_OHp7r5ePOQbInDMlZaMtRqYaTUVlLWcAdGyqEqmJWBCwOiS0IpQLSXjJSt1sRa8MJiTkoqKnmZXv96U5mOEEJveBpMSqAHcGBpe8YKWRCbwYg-Ouod1s_W2V35q_vpLwOUeUMGorvVqMDb8c0JiTGT5f3Fj3zaf1kMT-tR60tJGecIbumgITm_7AS0-iHQ</recordid><startdate>20060501</startdate><enddate>20060501</enddate><creator>KUCI, Zyrafete</creator><creator>SEITZ, Gabriele</creator><creator>KUCI, Selim</creator><creator>KREYENBERG, Hermann</creator><creator>SCHUMM, Michael</creator><creator>LANG, Peter</creator><creator>NIETHAMMER, Dietrich</creator><creator>HANDGRETINGER, Rupert</creator><creator>BRUCHELT, Gernot</creator><general>International Institute of Anticancer Research</general><scope>IQODW</scope><scope>CGR</scope><scope>CUY</scope><scope>CVF</scope><scope>ECM</scope><scope>EIF</scope><scope>NPM</scope><scope>7X8</scope></search><sort><creationdate>20060501</creationdate><title>Pitfalls in Detection of Contaminating Neuroblastoma Cells by Tyrosine Hydroxylase RT-PCR Due to Catecholamine-producing Hematopoietic Cells</title><author>KUCI, Zyrafete ; SEITZ, Gabriele ; KUCI, Selim ; KREYENBERG, Hermann ; SCHUMM, Michael ; LANG, Peter ; NIETHAMMER, Dietrich ; HANDGRETINGER, Rupert ; BRUCHELT, Gernot</author></sort><facets><frbrtype>5</frbrtype><frbrgroupid>cdi_FETCH-LOGICAL-h270t-ab955fbe5cfb3735f65e2b91845b9e022ecb423823b9956454b1d761c06243783</frbrgroupid><rsrctype>articles</rsrctype><prefilter>articles</prefilter><language>eng</language><creationdate>2006</creationdate><topic>Biological and medical sciences</topic><topic>Catecholamines - biosynthesis</topic><topic>Cell Line, Tumor</topic><topic>Dopamine beta-Hydroxylase - analysis</topic><topic>Dopamine beta-Hydroxylase - biosynthesis</topic><topic>Dopamine beta-Hydroxylase - genetics</topic><topic>Gene Expression Profiling</topic><topic>Hematopoietic Stem Cells - cytology</topic><topic>Hematopoietic Stem Cells - enzymology</topic><topic>Hematopoietic Stem Cells - metabolism</topic><topic>Humans</topic><topic>Leukocytes, Mononuclear - cytology</topic><topic>Leukocytes, Mononuclear - enzymology</topic><topic>Leukocytes, Mononuclear - metabolism</topic><topic>Medical sciences</topic><topic>Neuroblastoma - enzymology</topic><topic>Neuroblastoma - genetics</topic><topic>Neuroblastoma - pathology</topic><topic>Neurology</topic><topic>Reverse Transcriptase Polymerase Chain Reaction</topic><topic>Sensitivity and Specificity</topic><topic>Tumors</topic><topic>Tumors of the nervous system. Phacomatoses</topic><topic>Tyrosine 3-Monooxygenase - analysis</topic><topic>Tyrosine 3-Monooxygenase - biosynthesis</topic><topic>Tyrosine 3-Monooxygenase - genetics</topic><toplevel>peer_reviewed</toplevel><toplevel>online_resources</toplevel><creatorcontrib>KUCI, Zyrafete</creatorcontrib><creatorcontrib>SEITZ, Gabriele</creatorcontrib><creatorcontrib>KUCI, Selim</creatorcontrib><creatorcontrib>KREYENBERG, Hermann</creatorcontrib><creatorcontrib>SCHUMM, Michael</creatorcontrib><creatorcontrib>LANG, Peter</creatorcontrib><creatorcontrib>NIETHAMMER, Dietrich</creatorcontrib><creatorcontrib>HANDGRETINGER, Rupert</creatorcontrib><creatorcontrib>BRUCHELT, Gernot</creatorcontrib><collection>Pascal-Francis</collection><collection>Medline</collection><collection>MEDLINE</collection><collection>MEDLINE (Ovid)</collection><collection>MEDLINE</collection><collection>MEDLINE</collection><collection>PubMed</collection><collection>MEDLINE - Academic</collection><jtitle>Anticancer research</jtitle></facets><delivery><delcategory>Remote Search Resource</delcategory><fulltext>fulltext</fulltext></delivery><addata><au>KUCI, Zyrafete</au><au>SEITZ, Gabriele</au><au>KUCI, Selim</au><au>KREYENBERG, Hermann</au><au>SCHUMM, Michael</au><au>LANG, Peter</au><au>NIETHAMMER, Dietrich</au><au>HANDGRETINGER, Rupert</au><au>BRUCHELT, Gernot</au><format>journal</format><genre>article</genre><ristype>JOUR</ristype><atitle>Pitfalls in Detection of Contaminating Neuroblastoma Cells by Tyrosine Hydroxylase RT-PCR Due to Catecholamine-producing Hematopoietic Cells</atitle><jtitle>Anticancer research</jtitle><addtitle>Anticancer Res</addtitle><date>2006-05-01</date><risdate>2006</risdate><volume>26</volume><issue>3A</issue><spage>2075</spage><epage>2080</epage><pages>2075-2080</pages><issn>0250-7005</issn><eissn>1791-7530</eissn><abstract>Background: RT-PCR analysis of compounds of catecholamine metabolism (in particular tyrosine hydroxylase, TH) is widely used
for the detection of contaminating neuroblastoma cells in hematopoietic stem cell preparations. Due to reports in the literature
showing that hematopoietic cells are also able to produce catecholamines, we investigated whether TH-RT-PCR is really suitable
for this purpose. Materials and Methods: Besides neuroblastoma cells, mononuclear blood cells, apheresis preparations and
hematopoietic stem cells were used for single and nested RT-PCR. In addition to TH, the expressions of dopamine-β-hydroxylase
and noradrenaline transporter were analyzed. Results: Using single RT-PCR, a clear discrimination between neuroblastoma and
hematopoietic cells was possible. However, by using nested RT-PCR, the âneuroblastoma markersâ were also detected in a significant
percentage of non-mobilized mononuclear blood cells, in mononuclear blood cells of healthy donors mobilized with G-CSF, and
in highly purified CD34 + and CD133 + stem cells from healthy mobilized donors. Conclusion: Our results raise the question of whether the RT-PCR analysis of compounds
of catecholamine metabolism is suitable and selective enough to detect the contamination of hematopoietic stem cells by a
low number of neuroblastoma cells.</abstract><cop>Attiki</cop><pub>International Institute of Anticancer Research</pub><pmid>16827147</pmid><tpages>6</tpages></addata></record> |
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| subjects | Biological and medical sciences Catecholamines - biosynthesis Cell Line, Tumor Dopamine beta-Hydroxylase - analysis Dopamine beta-Hydroxylase - biosynthesis Dopamine beta-Hydroxylase - genetics Gene Expression Profiling Hematopoietic Stem Cells - cytology Hematopoietic Stem Cells - enzymology Hematopoietic Stem Cells - metabolism Humans Leukocytes, Mononuclear - cytology Leukocytes, Mononuclear - enzymology Leukocytes, Mononuclear - metabolism Medical sciences Neuroblastoma - enzymology Neuroblastoma - genetics Neuroblastoma - pathology Neurology Reverse Transcriptase Polymerase Chain Reaction Sensitivity and Specificity Tumors Tumors of the nervous system. Phacomatoses Tyrosine 3-Monooxygenase - analysis Tyrosine 3-Monooxygenase - biosynthesis Tyrosine 3-Monooxygenase - genetics |
| title | Pitfalls in Detection of Contaminating Neuroblastoma Cells by Tyrosine Hydroxylase RT-PCR Due to Catecholamine-producing Hematopoietic Cells |
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