PMID- 9404923
OWN - NLM
STAT- MEDLINE
DCOM- 19980217
LR  - 20151119
IS  - 0268-3369 (Print)
IS  - 0268-3369 (Linking)
VI  - 20
IP  - 10
DP  - 1997 Nov
TI  - Mobilization of hematopoietic progenitors in patients with chronic myeloid 
      leukemia.
PG  - 835-42
AB  - Although the mobilization and harvesting of Philadelphia chromosome-negative 
      progenitors has been proven feasible by a number of groups, it is not clear which 
      techniques should be used with regard to the monitoring of purging and evaluation 
      of stem cell yield. Therefore, we isolated CD34-positive cells from leukapheresis 
      samples of seven CML patients after induction therapy. These cells were analyzed 
      using the colony-forming unit granulocyte-macrophage (CFU-GM) and long-term 
      culture-initiating cell (LTCIC) assays. In addition, we analyzed frequency and 
      clonogenicity of single stem cells using the LTCIC assay at limiting dilution. 
      Individual colonies derived from these assays were subsequently analyzed for the 
      presence of the bcr-abl gene with interphase fluorescence in situ hybridization 
      (FISH) and for the presence of bcr-abl mRNA with RT-PCR. We compared these 
      results with the cytogenetic analysis of the leukapheresis material. Molecular 
      analysis of individual colonies correlated well with the results of cytogenetic 
      analysis. However, in nine out of 18 samples, cytogenetic analysis exclusively 
      demonstrated the presence of either Philadelphia chromosome-positive or -negative 
      cells whereas with the molecular analysis of individual colonies tumor 
      contamination or the presence of residual normal cells could be substantiated. We 
      concluded that molecular analysis of individual colonies should be employed to 
      further optimize induction protocols. With regard to stem cell mobilization we 
      demonstrated that about 67 CFU-GM colonies and clusters were generated by one 
      single LTCIC both for the CML samples and the samples obtained from patients with 
      non-hematologic malignancy. This number is important since until now most centres 
      use a number of four colonies and clusters generated by one LTCIC. Dividing the 
      CFU-GM output in the LTCIC assay by four to determine LTCIC frequency could thus 
      lead to an almost 20-fold overestimation of the LTCIC frequency. It is concluded 
      that stem cell frequency analysis is an important tool with regard to the 
      interpretation of mobilization protocols.
FAU - Thijsen, S F
AU  - Thijsen SF
AD  - Department of Hematology, Free University Hospital, Amsterdam, The Netherlands.
FAU - Schuurhuis, G J
AU  - Schuurhuis GJ
FAU - van Oostveen, J W
AU  - van Oostveen JW
FAU - Theijsmeijer, A P
AU  - Theijsmeijer AP
FAU - Niewint, A W
AU  - Niewint AW
FAU - Ossenkoppele, G J
AU  - Ossenkoppele GJ
LA  - eng
PT  - Journal Article
PL  - England
TA  - Bone Marrow Transplant
JT  - Bone marrow transplantation
JID - 8702459
RN  - 0 (RNA, Messenger)
RN  - 0 (RNA, Neoplasm)
RN  - 0 (Recombinant Proteins)
RN  - 04079A1RDZ (Cytarabine)
RN  - 143011-72-7 (Granulocyte Colony-Stimulating Factor)
RN  - EC 2.7.10.2 (Fusion Proteins, bcr-abl)
RN  - PVI5M0M1GW (Filgrastim)
RN  - ZS7284E0ZP (Daunorubicin)
SB  - IM
MH  - Adult
MH  - Clone Cells/cytology
MH  - Colony-Forming Units Assay
MH  - Cytarabine/pharmacology
MH  - Daunorubicin/pharmacology
MH  - Female
MH  - Filgrastim
MH  - Fusion Proteins, bcr-abl/genetics
MH  - Granulocyte Colony-Stimulating Factor/pharmacology
MH  - Hematopoietic Stem Cell Mobilization/*methods
MH  - Hematopoietic Stem Cell Transplantation/adverse effects
MH  - Humans
MH  - In Situ Hybridization, Fluorescence
MH  - Leukapheresis
MH  - Leukemia, Myelogenous, Chronic, BCR-ABL Positive/*blood/pathology/therapy
MH  - Male
MH  - Middle Aged
MH  - *Neoplastic Cells, Circulating
MH  - Neoplastic Stem Cells/chemistry/pathology
MH  - Polymerase Chain Reaction
MH  - RNA, Messenger/analysis
MH  - RNA, Neoplasm/analysis
MH  - Recombinant Proteins
MH  - *Tumor Stem Cell Assay
EDAT- 1997/12/24 00:00
MHDA- 1997/12/24 00:01
CRDT- 1997/12/24 00:00
PHST- 1997/12/24 00:00 [pubmed]
PHST- 1997/12/24 00:01 [medline]
PHST- 1997/12/24 00:00 [entrez]
AID - 10.1038/sj.bmt.1700991 [doi]
PST - ppublish
SO  - Bone Marrow Transplant. 1997 Nov;20(10):835-42. doi: 10.1038/sj.bmt.1700991.