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Evaluation of effects of Lingzhi mushroom (Ganoderma lucidum) on neural stem cells isolated from embryonic mouse brain (Mus musculus var. albino)

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Nguyen Thi Mai Dan, Truong Hai Nhung , Le Minh Dung, Nguyen Thuy Linh, Lam Thai Thanh and Pham Van Phuc

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Neural stem cells (NSCs) are employed as a promising tool for the replacement of damaged cells in several neurological disorders, including Parkinson’s, Alzheimer’s, Huntington’s diseases and spinal cord injury thanks to their self-renewal and pluripotency. However, due to the limitation in site-specific migration of endogenous stem cells, exogenous NSC sources, of which growth can be triggered by extrinsic factors, are taken advantage of. This study aims to evaluate the role of Ganoderma lucidum extract on neural stem cell proliferation as well as determine the concentration having the best growth stimulating effects. NSCs were isolated from 13.5 – 15.5 day embryonic mouse brain (Mus musculus var. albino) and cultured in DMEM/F-12 serum-free medium modified with B27, N2, heparin, EGF and FGF at 37oC, 5% CO 2 . Characteristics of candidate cells were evaluated by neurosphere assay, differentiation assay as well as the expression of Nestin, CD133 and Sox-1 by immunocytochemistry, flow cytometry and RT-PCR techniques, respectively. Those NSCs were then cultured in 96-well plate with Ganoderma lucidum extract at 100?g/ml, 500?g/ml and 1000?g/ml for evaluating mitogenic effects of such mushroom. The results revealed that 72% of cell sample was successfully cultured, cells isolated from 13.5-15.5 days post coitum fetus showed a high proliferation rate, isolated cells formed neurospheres, expressed neural stem cell markers and differentiated into GFAP positive cells. Ganoderma lucidum extract at 500?g/ml showed the best effects on neural stem cell growth.

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Ganoderma lucidum, neurosphere assay, neural stem cell markers, neural-colony forming cell assay, neurosphere size.