Quantifying mesenchymal stem cells in the mononuclear cell fraction of bone marrow samples obtained for cell therapy.

AIMS: The use of bone marrow mononuclear cells (BMMNCs) as a source of mesenchymal stem cells (MSCs) for therapy has recently attracted the attention of researchers because BMMNCs can be easily obtained and do not require in vitro expansion before their use. This study was designed to quantify the MSC population in bone marrow (BM) samples obtained for cell therapy using flow cytometry to detect the CD271 antigen. MATERIAL AND METHODS: Autologous BM was obtained by posterior superior iliac crest aspiration under topical anesthesia. Mononuclear cells isolated from the BM aspirate on a Ficoll density gradient were used to treat patients with pressure ulcer (n = 13) bone nonunions (n = 3) or diabetic foot ulcers (n = 5). RESULTS: Our flow cytometry data revealed a low percentage as well as a high variability among patients of CD271(+)CD45(-) cells (range, 0.0017 to 0.0201%). All cultured MSC adhered to plastic dishes showing a capacity to differentiate into adipogenic and osteogenic lineages. CONCLUSIONS: Our findings suggested that the success of cell therapy was independent of the number of MSCs present in the BM aspirate used for autologous cell therapy.

New serum-derived albumin scaffold seeded with adipose-derived stem cells and olfactory ensheathing cells used to treat spinal cord injured rats.

Recent advances in spinal cord injury (SCI) research and cell culture techniques and biomaterials predict promising new treatments for patients with SCI or other nerve injuries. Biomaterial scaffolds form a substrate within which cells are instructed to form a tissue in a controlled manner. This study was designed to assess axon regeneration and locomotor recovery in rats with spinal cord injury treated with a novel serum-derived albumin scaffold seeded with adipose derived stem cells (ADSCs) and olfactory ensheathing cells (OECs). OECs are considered promising candidates for the treatment of SCI, and ADSCs have the ability to differentiate into neural lineages. In vitro experiments revealed that ADSCs and OECs adhered to the scaffold, remained viable and expressed specific markers of their cell types when cultured in the scaffold. Rats treated with scaffold plus cells showed locomotor skills at several time points from 45 days post-injury that were improved over those recorded in control injured, untreated animals. Astrocytic scars and tissue regeneration, identified using histological and immunohistochemical techniques, revealed that although the scaffold itself appeared to play a significant role in reducing glial scar formation and filling of the lesion cavity with cells, the presence of ADSCs and OECs in the scaffold led to the appearance of cells expressing markers of neurons and axons at the injury site. Our findings point to the clinical feasibility of an albumin scaffold seeded with ADSCs and OECs as a treatment candidate for use in spinal cord injury repair studies.

Nefopam is more potent than carbamazepine for neuroprotection against veratridine in vitro and has anticonvulsant properties against both electrical and chemical stimulation.

Nefopam (NEF) is a known analgesic that has recently been shown to be effective in controlling both neuropathic pain and convulsions in rodents. In this study we compared nefopam to carbamazepine (CBZ), a reference antiepileptic drug (AED), for their ability to protect cerebellar neuronal cultures from neurodegeneration induced by veratridine (VTD). Furthermore, we tested nefopam for protection against both, maximal electroshock-induced seizures (MES), and isoniazid-induced seizures in mice. Both NEF and CBZ were effective in preventing both signs of excitotoxicity and neurodegeneration following exposure of cultures to 5 microM veratridine for 30 min and 24 h, respectively. Concentrations providing full neuroprotection were 500 microM CBZ and 50 microM NEF, while the concentration providing 50% neuroprotection was 200 microM for CBZ and 20 microM for NEF. Neither NEF nor CBZ reduced excitotoxicity following direct exposure of cultures to glutamate, but CBZ failed to reduce increases in intracellular calcium following stimulation of L-type voltage sensitive calcium channels. In vivo, NEF (20 mg/kg i.p.) significantly reduced MES and fully prevented MES-induced terminal clonus (TC). In comparison, NEF was significantly more effective than CBZ in preventing MES, although both drugs were equally effective against MES-induced TC. Furthermore, nefopam provided protection against isoniazid-induced seizures at doses similar to those protecting against MES.