Characterisation of multipotent stem cells from human peripheral blood using an improved protocol

This study was conducted to evaluate the multipotency of PB-derived MSCs (PB-MSCs) and effects of human PB-MSC transplantation on ectopic bone regeneration in nude mice. Ten millilitres of venous blood was collected from five healthy donors with heparin, and the red blood cells were removed using red blood cell lysis buffer. The cell suspension was cultured under normoxia culture and hypoxia culture conditions. The nonadherent cells were removed by changing 50% of the culture media every other day. The adherent cells were then passaged and subjected to in vitro multidifferentiation assays and in vivo ectopic bone formation assay. PB-MSCs were obtained and established in 60% of the samples within 1–2 weeks using the hypoxia culture protocol. Characterization assays indicated that cells cultured under hypoxia condition possessed potent multilineage differentiation capacities and expressed Nanog and Lgr5, as well as a series of MSC surface antigen markers (including CD29, CD90, CD105 and CD73). Newly formed bone tissues were confirmed by histological examinations in the ectopic bone formation assay. Thus, hypoxia is an effective and reliable method to improve the yield of PB-MSCs. The PB-MSCs established using our method showed potent multidifferentiation potentials. Lgr5 may be a potential biomarker for identification of a subpopulation of PB-MSCs which appeared to possess better osteogenic capacity. β-tricalcium phosphate scaffolds loaded with PB-MSCs is a promising os...
Source: Journal of Orthopaedic Translation - Category: Orthopaedics Source Type: research