Flow cytometry and fluorescence in situ hybridization to detect residual neuroblastoma cells in bone marrow

Mehmet Fatih Okcu; Rui Yu Wang; Carlos Bueso-Ramos; Wendy Schober; Douglas Weidner; Richard Andrassy; Martin Blakely; Heidi Russell; Alp Ozkan; John Kuttesch; Michael Andreeff; Ka Wah Chan; Joann Ater

doi:10.1002/pbc.20428

Flow cytometry and fluorescence in situ hybridization to detect residual neuroblastoma cells in bone marrow

Mehmet Fatih Okcu, Rui Yu Wang, Carlos Bueso-Ramos, Wendy Schober, Douglas Weidner, Richard Andrassy, Martin Blakely, Heidi Russell, Alp Ozkan, John Kuttesch, Michael Andreeff, Ka Wah Chan, Joann Ater

Research output: Contribution to journal › Article › peer-review

12 Scopus citations

Abstract

Background. In patients with neuroblastoma morphological assessment of BM for residual NB cells is not precise, particularly when the number of tumor cells is small. Procedure. To develop a sensitive and rapid method of detecting NB cells in BM, we assessed the efficiency of flow cytometry (FCM) using markers CD9, CD56, and CD45. The percent of CD9+/ CD56+/CD45- (NB phenotype) cells was determined by FCM in 41 samples (16 patients) at various time points. For confirmation fluorescence in situ hybridization (FISH) for 17q gain was performed. Results. Nineteen of the 22 (86%) samples that were negative by morphology were positive by FCM (>0.006% CD9+/ CD56+/CD45- cells). The longest time to complete the FCM study was 3 hr. In six FISH experiments the sorted CD9+/CD56+/CD45-population had a higher percentage of cells with 17q gain (11.5-95%) compared to a CD56-/ CD45+ internal control population (2-8%). Conclusions. Our preliminary results suggest that FCM determination of the percent of CD9+/ CD56+/CD45- cells is an effective method of rapidly detecting NB cells in BM.

Original language	English (US)
Pages (from-to)	787-795
Number of pages	9
Journal	Pediatric Blood and Cancer
Volume	45
Issue number	6
DOIs	https://doi.org/10.1002/pbc.20428
State	Published - Oct 2005

Keywords

Flow cytometry
Neuroblastoma
Residual disease

ASJC Scopus subject areas

Pediatrics, Perinatology, and Child Health
Hematology
Oncology

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10.1002/pbc.20428

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title = "Flow cytometry and fluorescence in situ hybridization to detect residual neuroblastoma cells in bone marrow",

abstract = "Background. In patients with neuroblastoma morphological assessment of BM for residual NB cells is not precise, particularly when the number of tumor cells is small. Procedure. To develop a sensitive and rapid method of detecting NB cells in BM, we assessed the efficiency of flow cytometry (FCM) using markers CD9, CD56, and CD45. The percent of CD9+/ CD56+/CD45- (NB phenotype) cells was determined by FCM in 41 samples (16 patients) at various time points. For confirmation fluorescence in situ hybridization (FISH) for 17q gain was performed. Results. Nineteen of the 22 (86%) samples that were negative by morphology were positive by FCM (>0.006% CD9+/ CD56+/CD45- cells). The longest time to complete the FCM study was 3 hr. In six FISH experiments the sorted CD9+/CD56+/CD45-population had a higher percentage of cells with 17q gain (11.5-95%) compared to a CD56-/ CD45+ internal control population (2-8%). Conclusions. Our preliminary results suggest that FCM determination of the percent of CD9+/ CD56+/CD45- cells is an effective method of rapidly detecting NB cells in BM.",

keywords = "Flow cytometry, Neuroblastoma, Residual disease",

author = "Okcu, {Mehmet Fatih} and Wang, {Rui Yu} and Carlos Bueso-Ramos and Wendy Schober and Douglas Weidner and Richard Andrassy and Martin Blakely and Heidi Russell and Alp Ozkan and John Kuttesch and Michael Andreeff and Chan, {Ka Wah} and Joann Ater",

year = "2005",

month = oct,

doi = "10.1002/pbc.20428",

language = "English (US)",

volume = "45",

pages = "787--795",

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T1 - Flow cytometry and fluorescence in situ hybridization to detect residual neuroblastoma cells in bone marrow

AU - Okcu, Mehmet Fatih

AU - Wang, Rui Yu

AU - Bueso-Ramos, Carlos

AU - Schober, Wendy

AU - Weidner, Douglas

AU - Andrassy, Richard

AU - Blakely, Martin

AU - Russell, Heidi

AU - Ozkan, Alp

AU - Kuttesch, John

AU - Andreeff, Michael

AU - Chan, Ka Wah

AU - Ater, Joann

PY - 2005/10

Y1 - 2005/10

N2 - Background. In patients with neuroblastoma morphological assessment of BM for residual NB cells is not precise, particularly when the number of tumor cells is small. Procedure. To develop a sensitive and rapid method of detecting NB cells in BM, we assessed the efficiency of flow cytometry (FCM) using markers CD9, CD56, and CD45. The percent of CD9+/ CD56+/CD45- (NB phenotype) cells was determined by FCM in 41 samples (16 patients) at various time points. For confirmation fluorescence in situ hybridization (FISH) for 17q gain was performed. Results. Nineteen of the 22 (86%) samples that were negative by morphology were positive by FCM (>0.006% CD9+/ CD56+/CD45- cells). The longest time to complete the FCM study was 3 hr. In six FISH experiments the sorted CD9+/CD56+/CD45-population had a higher percentage of cells with 17q gain (11.5-95%) compared to a CD56-/ CD45+ internal control population (2-8%). Conclusions. Our preliminary results suggest that FCM determination of the percent of CD9+/ CD56+/CD45- cells is an effective method of rapidly detecting NB cells in BM.

AB - Background. In patients with neuroblastoma morphological assessment of BM for residual NB cells is not precise, particularly when the number of tumor cells is small. Procedure. To develop a sensitive and rapid method of detecting NB cells in BM, we assessed the efficiency of flow cytometry (FCM) using markers CD9, CD56, and CD45. The percent of CD9+/ CD56+/CD45- (NB phenotype) cells was determined by FCM in 41 samples (16 patients) at various time points. For confirmation fluorescence in situ hybridization (FISH) for 17q gain was performed. Results. Nineteen of the 22 (86%) samples that were negative by morphology were positive by FCM (>0.006% CD9+/ CD56+/CD45- cells). The longest time to complete the FCM study was 3 hr. In six FISH experiments the sorted CD9+/CD56+/CD45-population had a higher percentage of cells with 17q gain (11.5-95%) compared to a CD56-/ CD45+ internal control population (2-8%). Conclusions. Our preliminary results suggest that FCM determination of the percent of CD9+/ CD56+/CD45- cells is an effective method of rapidly detecting NB cells in BM.

KW - Flow cytometry

KW - Neuroblastoma

KW - Residual disease

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Flow cytometry and fluorescence in situ hybridization to detect residual neuroblastoma cells in bone marrow

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