title: Characterisation of cryopreserved cells freshly isolated from human bone marrow.
abstract: Osteoblast progenitor cells (OBPCs) isolated from bone marrow have the ability to differentiate into osteoblasts and thus potential therapeutic use to tissue-engineer bone. In order for OBPCs to be available for clinical use a means of storing viable cells is necessary. The aim of this study was to determine whether a simple method of cryopreservation had an effect on osteogenic differentiation or growth of OBPCs isolated from fresh human bone marrow. Stro-1 was used to identify the isolated OBPCs. The osteoblastic potential of the marrow cells was confirmed as culture with osteogenic supplements (OS) significantly increased osteoblastic protein production (alkaline phosphatase (ALP), osteopontin and osteocalcin) compared with standard conditions (P less than 0.05). Ten further marrow aspirates were harvested; each was halved for either cryopreservation or control culture. Primary cultures from both populations formed colonies with recognised OBPC morphology. OS stimulated both cryopreserved and control populations to produce significantly more osteoblastic proteins (P less than 0.05) and there was no significant difference between the increase in osteogenic proteins when cultured with OS (P great than 0.2). The proliferation rate after 5 days in culture was not significantly affected by cryopreservation (P greater than 0.05). It has been suggested that OBPCs are immuno-privileged; so allogenic cells could be implanted into patients for tissue engineering bone without causing a hypersensitivity reaction. Our study demonstrates a method of storage, which allows OBPCs to be available for use without affecting osteoblastic potential or viability.
authors.affiliation: Department of Orthopaedics, Bonney House, St Marys Hospital, Praed Street, London, UK. philipparust@hotmail.com
authors.forename: Philippa A
authors.initials: PA
authors.lastname: Rust
authors.forename: Costas
authors.initials: C
authors.lastname: Tingerides
authors.forename: Stephen R
authors.initials: SR
authors.lastname: Cannon
authors.forename: Timothy W R
authors.initials: TW
authors.lastname: Briggs
authors.forename: Gordon W
authors.initials: GW
authors.lastname: Blunn
mesh_terms: D000469:Alkaline Phosphatase / Q000494:pharmacology; D001854:Bone Marrow Cells / Q000166:cytology* / Q000187:drug effects; D049109:Cell Proliferation / Q000187:drug effects; D002470:Cell Survival; D015925:Cryopreservation / Q000379:methods*; D003470:Culture Media; D006801:Humans; D010006:Osteoblasts / Q000166:cytology / Q000187:drug effects; D015675:Osteocalcin / Q000494:pharmacology; D010012:Osteogenesis / Q000187:drug effects; D053495:Osteopontin; D012795:Sialoglycoproteins / Q000494:pharmacology; D013234:Stem Cells / Q000166:cytology / Q000187:drug effects
chemical_list: D003470:Culture Media; C506895:SPP1 protein, human; D012795:Sialoglycoproteins; D015675:Osteocalcin; D053495:Osteopontin; D000469:Alkaline Phosphatase