Abstract

The seed extract of Mucuna gigantea was evaluated, its efficacy on the differentiation of mesenchymal stem cells (MSCs) into neural lineage. The collected seeds were extracted by using 1% aqueous acetic acid. According to High-performance liquid chromatography(HPLC) analysis, the average concentration of L-3,4-dihydroxyphenylalanine (L-DOPA) was 117.5±7.6 µg/mg of crude extract. The MSCs were cultured in neural induction medium with or without the extract. The induction medium was then replaced with the serum free medium. The induction process was monitored by phase contrast microscopy, immunocytochemistry and real-time polymerase chain reaction. After 14 days of cultivation, the cells cultured in the presence of the extract showed higher degree of nestin and β-III tubulin messenger ribonucleic acid (mRNA) expression than those cultured without the extract. The maximum gene expression was observed in cells cultured with 50 μg/ml of the extract. The induced MSCs showed a change in morphology to that of neuron-like cells. Immunocytochemical staining showed the expression of neural markers such as nestin and microtubule associated protein-2 (MAP-2) on day 7 and 28, respectively. This study suggests that the extract of M. gigantea, which contained L-DOPA and/or the other unidentified active compounds, could induce MSCs to express neural protein and gene markers.

Key words: Mucuna gigantean, L-DOPA, mesenchymal stem cells, neuron-like cells.

Abbreviation

 bFGF, Basic fibroblast growth factor; DMEM, dulbecco’s modified eagle medium; EGF, epidermal growth factor; FBS, fetal bovine serum; PD, Parkinson’s disease;MAP-2, microtubule associated protein-2; MSCs, mesenchymal stem cells; NM, neural induction medium; NM+MG, neural induction medium with M. gigantea; NSCs, neural stem