Immunocytochemistry Studies Using Microtubule-Associated Protein-2 (MAP-2) Markers on Neural Differentiation of Mesenchymal Stem Cells from Rat Bone Marrow

Abstract

Mesenchymal stem cells (MSCs) can differentiate into neurons, which can be used in cell therapy. Endogenous genes and neurotrophic factors such as nerve growth factor (NGF), brain-derived neurotrophic factor (BDNF), and neurotrophin-3 regulate neural stem cell proliferation and differentiation (NT-3). The differentiation of MSCs into neurons is characterized by the development of neurons that interact with each other to form rings or net-like structures. The markers of neurons in the immature stage are Nestin and Vimentin. While the markers for neurons at the mature stage are neuron-specific enolase (NSE), neuronalspecific nuclear protein (NeuN), and microtubule-associated protein-2 (MAP-2). This research aims to investigate the expression of MAP-2 on the differentiation of MSCs into mature neurons. MSCs were isolated from rat bone marrow of femur and tibia using flushing methods, then cultured in Minimum Essential Medium Eagle (MEM), 10% Fetal Bovine Serum (FBS), and 1% antibiotic-antimycotic. Neuron differentiation induction medium (MEM, 2% FBS, 1% insulin-like growth factor (N2), and NT-3 20, 25, and 30 ng/mL) was used to stimulate MSCs for 14 days (control induction medium without NT-3). The immunocytochemistry of MAP-2 was performed on day 14. All experiments were done triplicated. The data obtained is the average percentage of the number of MAP-2 positive cells at each concentration. SPSS statistical analysis with a one-way ANOVA test. The results showed a significant difference between the percentage of MAP-2 positive cells at NT-3 concentrations 20 (p < 0.05), 25 (p = 0,093), 30 ng/mL (p = 0,081), and the negative control. At concentrations of NT-3 20, 25, and 30 ng/mL the