THE EFFECTS OF β-MERCAPTOETHANOL ON NEUROGENIC DIFFERENTIATION ABILITY OF ADULT HUMAN STEM CELLS: A VISUAL CONFIRMATION

Durriyyah Sharifah HA*, Ramin K, Zaidah S, Nabilah HK, Siti Mariam AG, Shamsul Azlin AS, Wan Safwani WK, and Noor Hayaty AK

Abstract

THE EFFECTS OF β-MERCAPTOETHANOL ON NEUROGENIC DIFFERENTIATION ABILITY OF ADULT HUMAN STEM CELLS: A VISUAL CONFIRMATION Durriyyah Sharifah HA1*, Ramin K1, Zaidah S1, Nabilah HK1, Siti Mariam AG1, Shamsul Azlin AS1, Wan Safwani WK2, and Noor Hayaty AK3

1Institute of Biological Sciences, Faculty of Science, 2Faculty of Engineering, 3Faculty

of Dentistry, University of Malaya, 50603 Kuala Lumpur, Malaysia.

This preliminary study attempted in vitro protocols to isolate, culture, and induce human adipose tissue and teeth pulp derived mesenchymal stem cells (MSCs) into neuronal-like cells. MSCs were isolated from adipose tissue and teeth pulp, which were obtained with informed consent from UMMC and Dentistry clinic patients, respectively. Isolated MSCs were cultured and maintained in complete medium and incubated at 37ºC. Media was replaced every three days and the cells were subcultured on reaching 70%-80% confluency. Passage 5 MSCs were initially treated for 24 hours with different low concentrations of treatment doses (0.6mM, 0.9mM and 1.2mM) of β-mercaptoethanol (β- ME) to induce neuronal differentiation. After 24 hours treatment, media was changed and replaced with fresh serum free media. MSCs were then treated with higher β-ME concentrations (6mM, 9mM and 12mM) for 3 hours. Adipose and teeth pulp derived MSCs showed spindle-shape or fibroblast-like morphology. Upon exposure to β-ME, these MSCs displayed neuronal-like phenotype as detected under phase contrast microscopy. Morphologically neuronal-like cells were observed; each having extended processes attached to the cell body (soma). Thus, the findings propose the current protocol as suitable to differentiate human adipose tissue and teeth pulp MSCs into morphologically neuronal-like cells.

Keywords: neuronal human stem cells, β-mercaptoethanol, human adipose derived MSCs, human teeth pulp derived MSCs