E-ISSN 2218-6050 | ISSN 2226-4485
 

Research Article


In vitro model of ischemic stroke neurons from Macaca fascicularis

Ratih Rinendyaputri, Sela Septima Mariya, Frans Dany, Ariyani Noviantari, Dwi Budiono, Lisa Andriani Lienggonegoro, Hasta Handayani Idrus, Wireni Ayuningtyas, Rachmawati Noverina, Fathul Huda, Ahmad Faried.


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/NHPs 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 demonstrate of neurons as a 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/ICC with the marker β-tubulin. Expression of the apoptotic response was performed by 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.

Key words: Ischemic stroke, Macaca fascicularis, Neuron, Oxygen glucose deprivation, OGD


 
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