title: Limited engraftment capacity of bone marrow-derived mesenchymal cells following T-cell-depleted hematopoietic stem cell transplantation.
abstract: 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 x 10(4)/kg vs 1. 19 +/- 0.19 x 10(4)/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.
authors.affiliation: Department of Hematology, University of Parma, Parma, Italy.
authors.forename: D
authors.initials: D
authors.lastname: Cilloni
authors.forename: C
authors.initials: C
authors.lastname: Carlo-Stella
authors.forename: F
authors.initials: F
authors.lastname: Falzetti
authors.forename: G
authors.initials: G
authors.lastname: Sammarelli
authors.forename: E
authors.initials: E
authors.lastname: Regazzi
authors.forename: S
authors.initials: S
authors.lastname: Colla
authors.forename: V
authors.initials: V
authors.lastname: Rizzoli
authors.forename: F
authors.initials: F
authors.lastname: Aversa
authors.forename: M F
authors.initials: MF
authors.lastname: Martelli
authors.forename: A
authors.initials: A
authors.lastname: Tabilio
mesh_terms: D000328:Adult; D001854:Bone Marrow Cells / Q000166:cytology; D018929:Cell Culture Techniques; D002455:Cell Division / Q000235:genetics; D005260:Female; D006085:Graft Survival / Q000276:immunology*; D006410:Hematopoiesis / Q000235:genetics; D018380:Hematopoietic Stem Cell Transplantation / Q000379:methods* / Q000592:standards; D006650:Histocompatibility Testing; D006801:Humans; D017404:In Situ Hybridization, Fluorescence; D008212:Lymphocyte Depletion; D008297:Male; D008648:Mesoderm / Q000166:cytology*; D008875:Middle Aged; D009190:Myelodysplastic Syndromes / Q000628:therapy; D023461:Myeloid Progenitor Cells; D016133:Polymerase Chain Reaction; D012737:Sex Factors; D017154:Stromal Cells / Q000166:cytology; D014019:Tissue Donors; D018183:Transplantation Chimera / Q000235:genetics; D014184:Transplantation, Homologous / Q000379:methods