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BACKGROUND: How to induce endogenous neural stem cells (NSCs) to differentiate into needed neural cell types is a hot spot of current researches.OBJECTIVE: To compare differences between fetal bovine serum and Chinese herbal formula Naoluoxintong serum supplementation for inducing proliferation and differentiation in rat embryonic NSCs.DESIGN, TIME AND SETTING: An in vitro, serum pharmacology, comparative, observation study was performed from March to September in 2008 at the Laboratory of Neurodegenerative Diseases,College of Life Science in University of Science and Technology of China, the Key Laboratory Breeding Base of Acupuncture Foundation and Technology in Anhui University of Traditional Chinese Medicine, the Anhui Province Key Laboratory of R & D of Chinese Medicine, and at the Level 3 Laboratory of Molecular Biology of the State Administration of Traditional Chinese Medicine.MATERIALS: The Chinese herbal formula Naoluoxintong was produced by Radix Astragali, Radix Notoginseng, Rhizoma Chuanxiong, Scolopendra at Anhui University of Traditional Chinese Medicine. Mouse anti-rat nestin, glial fibrillary acidic protein, and galactocerebroside monoclonal antibodies, as well as rabbit anti-neuron-specific enolase polyclonal antibody were produced by Chemicon, Billerica, MA, USA. METHODS: Wistar rats aged 3 months were intragastrically infused with Naoluoxintong. Wistar rat embryonic NSCs (passage 8) were induced to proliferate and differentiate using 10% fetal bovine serum, 10% Naoluoxintong serum, and 10% rat serum.MAIN OUTCOME MEASURES: Phenotypic changes in cultured cells were detected using phase contrast microscopy, and cell proliferation and differentiation were observed using immunofluorescence staining.RESULTS: Proliferation and differentiation of embryonic NSCs was induced by three different types of blood serum. Although the differentiation time course with Naoluoxintong serum was later than with the other two methods, the differentiated cells were morphologically similar to mature neurons to a greater extent.CONCLUSION: Naoluoxintong serum supplementation induced differentiation of NSCs into neuronal-like cells and stimulated neuronal maturation.