Open AccessRapid clinical‐scale propagation of mesenchymal stem cells using cultures initiated with immunoselected bone marrow CD105 + cells
Author(s) -
Spiropoulos Antonia,
Theodosaki Maria,
Stefanaki Kalliopi,
Paterakis George,
Tzetis Maria,
Giannikou Krinio,
Petrakou Eftichia,
Dimopoulou Maria N.,
Papassotiriou Ioannis,
Roma Eleptheria S.,
Kanavakis Emmanuel,
Graphakos Stelios,
Goussetis Evgenios
Publication year - 2011
Publication title -
journal of cellular and molecular medicine
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.44
H-Index - 130
eISSN - 1582-4934
pISSN - 1582-1838
DOI - 10.1111/j.1582-4934.2010.01157.x
Subject(s) - mesenchymal stem cell , cd90 , bone marrow , haematopoiesis , stem cell , endoglin , biology , cell culture , context (archaeology) , microbiology and biotechnology , immunology , cd34 , paleontology , genetics
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.