In vitro model of ischemic stroke neurons from Macaca fascicularis.

Abstract

Background: Primary cells have the same heterogeneity and differentiation capacity that has potential as an in vitro ischemic stroke models. It is hoped that primary cells from non-human primates that have genetic similarities to humans can provide molecular information and become more accurate data for use in drug screening, especially stroke therapy. B27 is a supplement commonly used in neuronal cell cultures, but there are concerns that its effects will interfere with the neuroprotective processes of the drug candidates being tested.

Aim: This research will prove the demonstration of neurons as an ischemic stroke model and the effects of B27 in Macaca fascicularis (Mf) neurons as a model for ischemic stroke under oxygen glucose deprivation (OGD).

Methods: Neurons were obtained from a collection of biological materials collected during previous research. Neuronal validation was performed using immunocytochemistry with the marker β-tubulin. Expression of the apoptotic response was performed by real time- polymerase chain reaction/(RT-PCR) using Bax, BCL-2, caspase-9, and p53 gene markers. Characterization of neurons in terms of positive tβubulin markers and induction of OGD in neurons can be performed for 6 h to model ischemic stroke.

Results: This study showed that cultured neurons under OGD conditions can experience apoptosis, namely by increasing pro-apoptosis and decreasing anti-apoptosis. However, B27 supplementation increased the expression of anti-apoptotic Bcl-2 genes and decreased proapoptotic genes such as Bax, caspase 9, and p53.

Conclusion: Neuron culture from Mf can be used as an in vitro model of ischemic stroke, and B27 supplementation in neurons exerts neuroprotective effects on the induction of OGD.