title: Streamlining the generation of an osteogenic graft by 3D culture of unprocessed bone marrow on ceramic scaffolds.
abstract: Mesenchymal stromal cells are present in very low numbers in the bone marrow, necessitating their selective expansion on tissue culture plastic prior to their use in tissue-engineering applications. MSC expansion is laborious, time consuming, unphysiological and not economical, thus calling for automated bioreactor-based strategies. We and others have shown that osteogenic grafts can be cultured in bioreactors by seeding either 2D-expanded cells or by direct seeding of the mononuclear fraction of bone marrow. To further streamline this protocol, we assessed in this study the possibility of seeding the cells onto porous calcium phosphate ceramics directly from unprocessed bone marrow. Using predetermined volumes of bone marrow from multiple human donors with different nucleated cell counts, we were able to grow a confluent cell sheath on the scaffold surface in 3 weeks. Cells of stromal, endothelial and haematopoietic origin were detected, in contrast to grafts grown from 2D expanded cells, where only stromal cells could be seen. Upon implantation in nude mice, similar quantities of bone tissue were generated as compared to that obtained by using the conventional number of culture expanded cells from the same donor. We conclude that human osteogenic grafts can be efficiently prepared by direct seeding of cells from unprocessed bone marrow.
authors.affiliation: University of Twente, MIRA Institute for Biomedical Technology and Technical Medicine, Department of Tissue Regeneration, Enschede, The Netherlands. a.ganguly@tnw.utwente.nl
authors.forename: Anindita
authors.initials: A
authors.lastname: Chatterjea
authors.forename: Auke J S
authors.initials: AJ
authors.lastname: Renard
authors.forename: Christel
authors.initials: C
authors.lastname: Jolink
authors.forename: Clemens A
authors.initials: CA
authors.lastname: van Blitterswijk
authors.forename: Jan
authors.initials: J
authors.lastname: de Boer
mesh_terms: D000368:Aged; D000369:Aged, 80 and over; D000818:Animals; D001853:Bone Marrow / Q000187:drug effects*; D002452:Cell Count; D049109:Cell Proliferation / Q000187:drug effects; D002470:Cell Survival / Q000187:drug effects; D002516:Ceramics / Q000494:pharmacology*; D005260:Female; D006801:Humans; D008297:Male; D059630:Mesenchymal Stem Cells / Q000166:cytology / Q000187:drug effects / Q000378:metabolism; D051379:Mice; D016513:Mice, SCID; D008875:Middle Aged; D010012:Osteogenesis / Q000187:drug effects*; D016062:Porosity / Q000187:drug effects; D046509:Tissue Culture Techniques / Q000379:methods*; D014019:Tissue Donors; D023822:Tissue Engineering / Q000379:methods*; D054457:Tissue Scaffolds / Q000737:chemistry*