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Blood, 15 November 2000, Vol. 96, No. 10, pp. 3637-3643
TRANSPLANTATION
Limited engraftment capacity of bone marrow-derived mesenchymal
cells following T-cell-depleted hematopoietic stem cell
transplantation
Daniela Cilloni,
Carmelo Carlo-Stella,
Franca Falzetti,
Gabriella Sammarelli,
Ester Regazzi,
Simona Colla,
Vittorio Rizzoli,
Franco Aversa,
Massimo F. Martelli, and
Antonio Tabilio
From the Department of Hematology, University
of Parma, Parma, Italy; the Department of Internal Medicine, University
of Torino, Torino, Italy; and the Department of Hematology, University
of Perugia, Perugia, Italy.
The engraftment capacity of bone marrow-derived mesenchymal
cells was investigated in 41 patients who had received a
sex-mismatched, T-cell-depleted allograft from human leukocyte antigen
(HLA)-matched or -mismatched family donors. Polymerase chain reaction
(PCR) analysis of the human androgen receptor (HUMARA) or the
amelogenin genes was used to detect donor-derived mesenchymal cells.
Only 14 marrow samples (34%) from 41 consenting patients generated a
marrow stromal layer adequate for PCR analysis. Monocyte-macrophage contamination of marrow stromal layers was reduced below the
levels of sensitivity of HUMARA and amelogenin assays (5% and
3%, respectively) by repeated trypsinizations and treatment with the
leucyl-leucine (leu-leu) methyl ester. Patients who received allografts
from 12 female donors were analyzed by means of the HUMARA assay, and in 5 of 12 cases a partial female origin of stromal cells was demonstrated. Two patients who received allografts from male donors were analyzed by amplifying the amelogenin gene, and in both cases a
partial male origin of stromal cells was shown. Fluorescent in situ
hybridization analysis using a Y probe confirmed the results of PCR
analysis and demonstrated in 2 cases the existence of a mixed
chimerism at the stromal cell level. There was no statistical difference detected between the dose of fibroblast progenitors (colony-forming unit-F [CFU-F]) infused to patients with
donor- or host-derived stromal cells
(1.18 ± 0.13 × 104/kg vs
1.19 ± 0.19 × 104/kg; P  .97).
In conclusion, marrow stromal progenitors reinfused in patients
receiving a T-cell-depleted allograft have a limited capacity of
reconstituting marrow mesenchymal cells.
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