Different studies have provided evidence that implantation of bone-marrow mononuclear cells (BM-MNC) into ischaemic limbs can improve tissue vascularization. Based on these results we performed a pilot study in patients with critical lower limb ischaemia to assess efficacy and safety of implantation of autologous BM-MNC. The amount and efficacy of BM-MNC purified either by an automated method or by a manual procedure were compared. Twelve patients with severe unilateral lower limb ischaemia were entered into this study. They were randomly assigned to be injected with BM-MNC sorted on a blood cell separator or isolated by density gradient on Ficoll-Hypaque. BM-MNC were implanted into the ischaemic legs. Patients were monitored with resting ankle-brachial pressure index (ABI), arterial oxygen saturation (SaO(2)), pain-free walking time and rest pain scale evaluation. The automated and manual methods used for mononuclear cell separation gave results not significantly different. Monitored variables improved in both groups. Improvement of ischaemic condition persisted during 24 weeks follow-up. Limb salvage was achieved in five cases. Our results indicate that BM-MNC implantation into ischaemic limbs is a practical, safe and effective method that may significantly contribute to the management of patients with limb ischaemia. The Ficoll method is a simple and effective procedure for BM-MNC concentration that may be useful, mainly in hospitals without sophisticated facilities.