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Quantitative analysis of cobblestone area-forming cells in bone marrow of
patients with aplastic anemia by limiting dilution assay
H Schrezenmeier, M Jenal, F Herrmann, H Heimpel and A Raghavachar
Department of Medicine III, University of Ulm, Germany.
In the past, the analysis of primitive human hematopoietic progenitor cells
with repopulating activity was limited by lack of appropriate in vitro
assay systems. It was recently shown that cobblestone area- forming cells
(CAFC) giving rise to cobblestone areas after 5 weeks in long-term marrow
cultures (LTMC) represent a population of pluripotent progenitor cells with
long-term marrow-repopulating activity. We have used a microtiter limiting
dilution-type human LTMC system to quantitate the frequency of CAFC (week
5) in aplastic anemia (AA). In bone marrow mononuclear cells (BM-MNC) of
healthy donors (n = 36) we observed a mean frequency of 84.4 CAFC per 10(5)
BM-MNC (95% confidence interval limits, 66.4 to 102.4). The mean frequency
of CAFC in BM of 31 AA patients was 6.6 per 10(5) BM-MNC (95% confidence
interval limits, 5.3 to 7.9; n = 47). This frequency is significantly lower
as compared with controls (P < .0001). The frequency of CAFC was reduced
not only in pancytopenic AA patients (6.2 per 10(5) BM-MNC; P < .0001 v
control), but also in patients in remission after immunosuppression (7.6; P
< .0001 v control; P = .1 v pancytopenic AA patients). The CAFC
frequency did not correlate with the severity or duration of the disease
and did not predict response to immunosuppressive treatment. In summary,
the frequency of primitive hematopoietic progenitor cells, as measured by
the CAFC assay, is significantly reduced in AA. CAFC remain severely
reduced even after hematologic recovery after immunosuppressive treatment.
The low frequency of CAFC in remission patients is in keeping with other
data pointing to a persisting defect of hematopoiesis in patients in
remission after immunosuppressive treatment.
Volume 88,
Issue 12,
pp. 4474-4480,
12/15/1996
Copyright © 1996 by The American Society of Hematology
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