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


METHODS
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.
Okcu
Mehmet Fatih
MF
Division of Pediatrics, The University of Texas M.D. Anderson Cancer Center, Houston, TX 77030, USA.
Wang
Rui-Yu
RY
Bueso-Ramos
Carlos
C
Schober
Wendy
W
Weidner
Douglas
D
Andrassy
Richard
R
Blakely
Martin
M
Russell
Heidi
H
Ozkan
Alp
A
Kuttesch
John
J
Andreeff
Michael
M
Chan
Ka Wah
KW
Ater
Joann
J
D000293:Adolescent; D000328:Adult; D015703:Antigens, CD / Q000032:analysis; D001853:Bone Marrow / Q000473:pathology*; D001856:Bone Marrow Examination; D002648:Child; D002675:Child, Preschool; D002869:Chromosome Aberrations; D002886:Chromosomes, Human, Pair 17; D005260:Female; D005434:Flow Cytometry / Q000379:methods / Q000592:standards; D006801:Humans; D016130:Immunophenotyping; D017404:In Situ Hybridization, Fluorescence / Q000379:methods / Q000592:standards; D007223:Infant; D008297:Male; D018365:Neoplasm, Residual / Q000175:diagnosis*; D009447:Neuroblastoma / Q000175:diagnosis / Q000473:pathology*; D012680:Sensitivity and Specificity
D015703:Antigens, CD