title: Rapid clinical-scale propagation of mesenchymal stem cells using cultures initiated with immunoselected bone marrow CD105+ cells.
abstract: Current clinical protocols used for isolation and purification of mesenchymal stem cells (MSC) are based on long-term cultures starting with bone marrow (BM) mononuclear cells. Using a commercially available immunoselection kit for enrichment of MSC, we investigated whether culture of enriched BM-CD105(+) cells could provide an adequate number of pure MSC in a short time for clinical use in the context of graft versus host disease and graft failure/rejection. We isolated a mean of 5.4 × 10(5) ± 0.9 × 10(5) CD105(+) cells from 10 small volume (10-25 ml) BM samples achieving an enrichment >100-fold in MSC. Seeding 2 × 10(3) immunoselected cells/cm(2) we were able to produce 2.5 × 10(8) ± 0.7 × 10(8) MSC from cultures with autologous serum enriched medium within 3 weeks. Neither haematopoietic nor endothelial cells were detectable even in the primary culture cell product. Expanded cells fulfilled both phenotypic and functional current criteria for MSC; they were CD29(+), CD90(+), CD73(+), CD105(+), CD45(-); they suppressed allogeneic T-cell reaction in mixed lymphocyte cultures and retained in vitro differentiation potential. Moreover, comparative genomic hybridization analysis revealed chromosomal stability of the cultured MSC. Our data indicate that adequate numbers of pure MSC suitable for clinical applications can be generated within a short time using enriched BM-CD105(+) cells.
authors: Stem Cell Transplant Unit, 'Aghia Sophia' Children's Hospital, Athens, Greece.
authors: Antonia
authors: A
authors: Spiropoulos
authors: Maria
authors: M
authors: Theodosaki
authors: Kalliopi
authors: K
authors: Stefanaki
authors: George
authors: G
authors: Paterakis
authors: Maria
authors: M
authors: Tzetis
authors: Krinio
authors: K
authors: Giannikou
authors: Eftichia
authors: E
authors: Petrakou
authors: Maria N
authors: MN
authors: Dimopoulou
authors: Ioannis
authors: I
authors: Papassotiriou
authors: Eleptheria S
authors: ES
authors: Roma
authors: Emmanuel
authors: E
authors: Kanavakis
authors: Stelios
authors: S
authors: Graphakos
authors: Evgenios
authors: E
authors: Goussetis
mesh_terms: D000328:Adult; D015703:Antigens, CD / Q000378:metabolism*; D001854:Bone Marrow Cells / Q000166:cytology* / Q000378:metabolism*; D018929:Cell Culture Techniques / Q000379:methods*; D002454:Cell Differentiation; D049109:Cell Proliferation; D002469:Cell Separation; D002478:Cells, Cultured; D002648:Child; D055028:Comparative Genomic Hybridization; D000071063:Endoglin; D006801:Humans; D016130:Immunophenotyping / Q000379:methods*; D007959:Lymphocyte Culture Test, Mixed; D008214:Lymphocytes / Q000166:cytology; D059630:Mesenchymal Stem Cells / Q000166:cytology* / Q000378:metabolism; D011956:Receptors, Cell Surface / Q000378:metabolism*; D055815:Young Adult
chemical_list: D015703:Antigens, CD; C064206:ENG protein, human; D000071063:Endoglin; D011956:Receptors, Cell Surface