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

DOI

10.2991/978-94-6463-112-8_6How to use a DOI?

Keywords

Differentiation; Immunocytochemistry; Microtubule-Associated Protein-2; Mesenchymal Stem Cells; Neuron

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), neuronal-specific 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 results showed a significant difference in the percentage of MAP-2 positive cells 13.82 ± 1.61%, 11.50 ± 1.38%, 11.34 ± 1.29% versus the control 8,24 ± 0.70%, respectively. The study confirms that MSCs can differentiate into mature neurons and can be confirmed by the MAP-2 marker and 20 ng/mL NT-3 could be used as a single inducer of MSCs differentiation into the mature neurons, indicated by the presence of MAP-2 marker.

Copyright

© 2023 The Author(s)

Open Access

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TY  - CONF
AU  - Ariyani Noviantari
AU  - Radiana D. Antarianto
AU  - Lutfah Rif’ati
AU  - Ratih Rinendyaputri
AU  - Uly A. Nikmah
AU  - Lisa A. Lienggonegoro
AU  - Masagus Zainuri
AU  - Frans Dany
PY  - 2023
DA  - 2023/03/01
TI  - Immunocytochemistry Studies Using Microtubule-Associated Protein-2 (MAP-2) Markers on Neural Differentiation of Mesenchymal Stem Cells from Rat Bone Marrow
BT  - Proceedings of the 1st International Conference for Health Research – BRIN (ICHR 2022)
PB  - Atlantis Press
SP  - 51
EP  - 64
SN  - 2468-5739
UR  - https://doi.org/10.2991/978-94-6463-112-8_6
DO  - 10.2991/978-94-6463-112-8_6
ID  - Noviantari2023
ER  -