Gene expression in differentiation of rat bone marrow-derived mesenchymal stem cells into neurocyte likt cells induced by salvia mitiorrhiza / 中国组织工程研究

ABSTRACT

BACKGROUND:It has been reported that mesenchymal stromal cells(MSCs)are capable of differentiating into cells of multilineage.Different methods and reagents have been used to induce the differentiation of MSCs,but most inducing systems contain serum and cytokines.OBJECTIVE:To investigate the gene expression of mash-1 and ngn-1 in differentiation of SD rat bone marrow-derived mesenchymal stem cells induced by salvia mitiorrhiza. DESIGN:Controlled experiment in vitro with repeated observation and measurement based on cells.SETTING:Department of Anatomy,School of Preclinical and Forensic Medicine,Sichuan University. MATERIALSThis study was performed in the Department of Anatomy,School of Preclinical and Forensic Medicine,Sichuan University from October 2004 to December 2005.SD male rats weighing 160-200 g were purchased from the Animal Center of Sichuan University.The experimental animals were disposed according to ethical criteria.Parenteral solution of salvia mitiorrhiza purchased from Tianyang Medicine Company Limited of Anhui(batch number20050411).METHODS:The nucleated cells were separated from rat bone marrow through gradient centrifugation and cell adherent method,and then MSCs differentiated into neurcyte-like cells induced by salvia mitiorrhiza.Cells in the control group were cultured with salvia mitiorrhiza-free serum-free culture media.The expression of neuron specific enolase(NSE)and glial fibrillary acidic protein(GFAP)were detected by immunohistochemistry.RT-PCR was used to detect the mRNA of mash-1 and ngn-1. MAIN OUTCOME MEASURES:①mash-1 and ngn-1 expressions were detect by the RT-PCR method.②NSE and GFAP expressions were detected by immunohistochemistry.RESULTS:①The MSCs were well adherent to walls.Most of the cells transformed in dipolar-like or multipolar-like.Axon-like or dentrite-like process was developed and these processes synapse with each other.② RT-PCR showed that ngn-1 and mash-1 mRNA were negative before induction,but positive after induction.③Immunohistochemistry indicated that NSE and GFAP expressions were positive after induction but negative in the control group. CONCLUSION:MSCs can be induced to differentiate into neurocyte-like cells by salvia mitiorrhiza.