title: Mesenchymal Stromal Cell Culture and Delivery in Autologous Conditions: A Smart Approach for Orthopedic Applications.
abstract: Human Mesenchymal Stromal Cells (hMSCs) are cultured in vitro with different media. Limits on their use in clinical settings, however, mainly depend on potential biohazard and inflammation risks exerted by xenogeneic nutrients for their culture. Human derivatives or recombinant materials are the first choice candidates to reduce these reactions. Therefore, culture supplements and materials of autologous origin represent the best nutrients and the safest products. Here, we describe a new protocol for the isolation and culture of bone marrow hMSCs in autologous conditions - namely, patient-derived serum as a supplement for the culture medium and fibrin as a scaffold for hMSC administration. Indeed, hMSC/fibrin clot constructs could be extremely useful for several clinical applications. In particular, we focus on their use in orthopedic surgery, where the fibrin clot derived from the donor's own blood allowed effective cell delivery and nutrient/waste exchanges. To ensure optimal safety conditions, it is of the utmost importance to avoid the risks of hMSC transformation and tissue overgrowth. For these reasons, the approach described in this paper also indicates a minimally ex vivo hMSC expansion, to reduce cell senescence and morphologic changes, and short-term osteo-differentiation before implantation, to induce osteogenic lineage specification, thus decreasing the risk of subsequent uncontrolled proliferation.
authors.affiliation: Dept. of Clinical and Experimental Medicine, University of Pisa; OtoLab, Azienda Ospedaliero-Universitaria Pisana (AOUP); l.trombi@yahoo.it.
authors.forename: Luisa
authors.initials: L
authors.lastname: Trombi
authors.affiliation: OtoLab, Azienda Ospedaliero-Universitaria Pisana (AOUP); Dept. of Civil and Industrial Engineering, University of Pisa.
authors.forename: Serena
authors.initials: S
authors.lastname: Danti
authors.affiliation: Immunohematology Operative Unit, Azienda Ospedaliero-Universitaria Pisana (AOUP).
authors.forename: Sara
authors.initials: S
authors.lastname: Savelli
authors.affiliation: Dept. of Clinical and Experimental Medicine, University of Pisa.
authors.forename: Stefania
authors.initials: S
authors.lastname: Moscato
authors.affiliation: OtoLab, Azienda Ospedaliero-Universitaria Pisana (AOUP); Dept. Of Surgical, Medical, Molecular Pathology and Emergency Medicine, University of Pisa.
authors.forename: Delfo
authors.initials: D
authors.lastname: D'Alessandro
authors.affiliation: OtoLab, Azienda Ospedaliero-Universitaria Pisana (AOUP).
authors.forename: Claudio
authors.initials: C
authors.lastname: Ricci
authors.affiliation: II Orthopedic and Traumatologic Clinic, Azienda Ospedaliero-Universitaria Pisana (AOUP).
authors.forename: Stefano
authors.initials: S
authors.lastname: Giannotti
authors.affiliation: Dept. of Clinical and Experimental Medicine, University of Pisa.
authors.forename: Mario
authors.initials: M
authors.lastname: Petrini
mesh_terms: D018929:Cell Culture Techniques; D002454:Cell Differentiation; D049109:Cell Proliferation; D002478:Cells, Cultured; D003470:Culture Media; D005337:Fibrin / Q000737:chemistry; D006801:Humans; D045164:Mesenchymal Stem Cell Transplantation; D059630:Mesenchymal Stem Cells / Q000166:cytology*; D010012:Osteogenesis; D044967:Serum / Q000737:chemistry; D054457:Tissue Scaffolds
chemical_list: D003470:Culture Media; D005337:Fibrin