TF-1 Cell Proliferation Assay for estimating Bioactivity of NeuroEPO
Keywords:
Palabras claves; NeuroEPO; neurogeneración; neuroprotección; bioensayoAbstract
Considerable upregulation of EPO/EPOR in brain occurs for instance upon brain injury, which stimulated researchers early to explore the neuroprotective and neuroregenerative potential of this growth factor. A new nasal formulation containing a non-hematopoietic recombinant EPO (NeuroEPO) has demonstrated neuroprotective actions in different preclinical models. NeuroEPO has a neuroprotective effect by positive modulation of the expression of antioxidant enzymes, a neurotrophic activity, an antiapoptotic action and an anti-inflammatory effect. However, the biological activity of the product has not been evaluated in in vitro models. In this work we aim to demonstrate that the TF1 cell line is an appropriate biomodel to evaluate the biological activity of the molecule. To do this, the expression of EPOR in the cell line was evaluated and then subsequently the cell proliferation in the presence of cytokines (GM-CSF and NeuroEPO) using trypan blue staining and the proliferation index with the Ki67 protein by flow cytometry were determined. In addition, cell proliferation was also measured using a colorimetric assay with Alamar Blue. The TF1 cell line was shown to express EPOR and it is a suitable in vitro model to evaluate the biological activity of NeuroEPO.
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