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Induction of Adult Rat Bone Marrow Stem Cells into 5-Hydroxytryptamine-sensitive Neurons in Vitro
Chang Shu, Ting-Yu Li*, Connie-Hau-Yan Wong1, Miu-Ching Lau1, Lai-Ling Tsang1, Hu Zhu1, Yu-Lin Gou1, Yiu-Wa Chung1, Hsiao-Chang Chan1
Children's Hospital, Chongqing University of Medical Sciences, Chongqing 400014, China; 1Epithelial Cell Biology Research Center, Department of Physiology, Faculty of Medicine, The Chinese University of Hong Kong, Hong Kong, China
Abstract: We successfully induced adult rat bone marrow stromal cells ( rMSCs) into phenotypic and functional neurons. To induce neuronal differentiation， rMSCs of passage 13_25 were grown in pre-induction media containing 10_7 mol/l all-trans-retinoic acid and 10 ng/ml basic fibroblast growth factor. After the pre-induction media were removed， the cells were transferred to serum free modified neuronal induction media. Within 3 h after induction， responsive cells assumed neuronal morphological characteristics. Neuron-specific enolase could be detected in 5 h and a (92.4±6.9)% of neuron-specific enolase positive cells was determined. Neurofilament H was also detected， with a (93.9±5.2)% of Neurofilament H positive cells observed after 24 h. Intracellular Ca2+ response to a ubiquitous neurotransmitter， 5-hydroxytryptamine， was monitored since both 5-hydroxytryptamine and Ca2+ are well known for their regulatory roles in neuronal proliferation， migration and differentiation. The results showed that the sensitivity of intracellular Ca2+ changes to 5-hydroxytryptamine increased with time with distinct Ca2+ spike similar to that in neurons observed 24 h after induction. Expression of 5-hydroxytryptamine1A receptor was weakly detected in stem cells， but with strong intensity observed in differentiated neurons. The present study has demonstrated， with well defined culture conditions， that adult rMSCs can be induced to phenotypic and functional neurons in vitro.
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