RESUMEN
BACKGROUND: Therapy with diverse cell types has been proposed to regenerate spinal cord injuries seeking to minimize the consequences for the lives of chronic patients. The types considered are: mononuclear and mesenchymal adult stem cells, embryonic stem cells, and Schwann cells. MATERIALS AND METHODS: Ninety male Wistar rats that underwent spinal cord contusion injury (NYU Impactor) were followed with the Basso, Beattie, and Bresnahan (BBB) locomotor rating scale for 14 days. Animals with scores < or = 16 were randomly divided into 2 groups: control (vehicle) versus cell therapy group. The mononuclear fraction (CD45(+)/CD34(-)) obtained by puncture-aspiration of the bone marrow was isolated by a density gradient (d = 1.077). The parenchymal cell infusion was performed using a syringe (100 U/1 mL) with a 30G1/2 needle. The animals were followed for 10 days before euthanasia. Statistical analyses comparing groups were performed by the Mann-Whitney test and group comparisons by the Wilcoxon test. RESULTS: Among 90 injured rats, 65 (72.2%) survived, including 44 whose scores were < or = 16. Eleven animals finished the study in the control group (64.7%) and 17 in the therapy group (80.9%). The statistical analyses did not demonstrate significance (P > .05) for either test. CONCLUSION: Mononuclear adult stem cell therapy was not demonstrated to be functionally effective for chronic spinal cord injury.
Asunto(s)
Trasplante de Médula Ósea/métodos , Traumatismos de la Médula Espinal/cirugía , Trasplante de Células Madre/métodos , Animales , Modelos Animales de Enfermedad , Masculino , Actividad Motora , Regeneración Nerviosa , Ratas , Ratas Wistar , Traumatismos de la Médula Espinal/fisiopatología , Resultado del TratamientoRESUMEN
The product generated by skeletal muscle and bone marrow mesenchymal stem cell cocultures has been demonstrated to improve the functional outcomes after cell therapy in postinfarction or Chagas myocardiopathy. This coculture method allows cell interactions in vitro, diminishing the operational costs of the culture/expansion as well as leading to angiogenesis and myogenesis for regeneration of the injured heart. Flow cytometric analysis may better characterize the cellular types in this model. Our objective was to use flow cytometry to analyze the immunophenotype expressed in this coculture model. The coculture was performed in accordance with Carvalho for 21 days. Flow cytometry was performed before and after coculture to characterize the immunophenotypic profile of cellular subsets, namely, the surface markers CD31, CD34, CD44H, CD45, CD49d, CD54, CD73, CD90, CD105, CD106, Myo-D, M-cadherin, and Connexin-43. Statistics were performed by the nonparametric Friedman test (P < .05) with post-hoc analysis by the nonparametric Wilcoxon test (P < or = .017, Bonferroni correction). The results demonstrated statistical significance for CD45(+) in 89.49% of mononuclear cells, 3.58% in skeletal muscle cells, and 4.74% among cocultured cells (P = .0094); and CD90(+) in 36.18% of mononuclear cells, 6.01% in skeletal muscle cells, and 48.94% among cocultured cells (P = .0420). The cocultured cells expressed the markers CD73(++), CD90(+++), CD45(-), CD34(+), CD105(-/+), CD106(-/+), M-cadherin(-/+), and Connexin-43(-/+). In conclusion, flow cytometric analysis showed a heterogeneous adherent cell population in this coculture model.