Induction of Human Umbilical Cord Blood Mesenchymal Stem Cells into Nerve-Like Cells by Salvia Miltiorrhiza.

Salvia miltiorrhiza is a traditional Chinese herb with thousands of years history. Composite salvia injection (CSI), a widely used administration in China nowadays, consists of Salvia miltiorrhiza, notoginseng, radix and borneo. It can promote blood flow, but also regulate qi to alleviate pain. So it is frequently used for angina, coronary heart disease and chest discomfort. Recently, some laboratory studies show that CSI may have the effect of cell induction.. In this study, we aim to investigate its neural induction.

Objective: To investigate the differentiation-inducing effects of salvia miltiorrhiza on umbilical cord blood mesenchymal stem cells.

Methods: The immunophenotypes of human umbilical cord blood-derived mesenchymal stem cells(MSCs) were determined by flow-cytometry. Salvia miltiorrhiza was adapted to induce the differentiation of the primary cultured cells of umbilical cord blood and MSCs. The results were compared with nerve growth factor(NGF) and ganglioside(GM1). The differentiated and undifferentiated cells were identified with immunocytochemistry.

Results: In primary culture, certain cells proliferated and formed cell clusters. These cells were either fibroblast-like or round in shape with various sizes. The positive rates for MSCs phenotypes CD29, CD44, CD59 of primary culture cells were 10.7%,37.27% and 66.67%, respectively. Salvia miltiorrhiza could induce the primary cultured cells of umbilical cord blood to express neural phenotypes. After passage, the MSCs could be maintained in vitro for extended periods with stable population doubling. The positive rates for CD29, CD44, CD59 of human umbilical cord blood-derived MSCs were 40.2%,70.5% and 95.4%, respectively. Salvia miltiorrhiza could induce the MSCs to express three kinds of proteins: nestin(the marker of neuronal precursor stem cells), β-TubulinIII and neurofilament (NF) (the marker of neuronal phenotypes) and glial fibrillary acidic protein(GFAP, the marker of astrocyte). Besides, NGF and GM1 could also induce the same MSCs to nerve-like cell, which could express the same markers: β-TubulinIII, NF and GFAP. In comparison NGF and GM1, the induction effect of salvia miltiorrhiza demonstrated the higher rate of expressing neuron phenotypes at a faster speed.

Conclusion: The human MSCs could be isolated from human umbilical cord blood. Salvia miltiorrhiza can induce the MSCs to differentiate into nerve-like cells, therefore, might act as a potential drug for neural induction.