The Expression Of Nestin-m RNA In Umbilical Mesenchymal Stem Cells

Objective To observe the expression of umbilical cord mesenchymal stem cells cultured charge in nerve cells of nestin nestin-m RNA. Methods charge separation between the umbilical cord mesenchymal stem cells were cultured and purified. Based on the health of mothers willing to donate umbilical cord after birth, isolated single cells, count cells from the umbilical cord by enzymatic digestion, the detection of activity by flow cytometry. Extraction of primary, second-generation cellular m RNA, which express nestin-m RNA was detected by real-time PCR method. Result Cells isolated from umbilical cord adherent growth in good condition, cell viability was 96.63% ~ 99.70%. Primary culture experiments, the fresh umbilical cord after treatment were observed visible freshly isolated MSCs round cell body, where a large part of the growth of fusiform cells, the third day, the cells begin adherent cells were spindle or triangles, smaller, scattered throughout the dish, a small amount of visible debris suspension cultured cells, the cells were cultured to grow a lot faster than the slow increase in the first seven days of spindle cells, cell clusters tend to grow; after passaging cells and primary similar cell morphology, growth accelerated;individual nerve-like cells were grown. Cell culture at passage P5-generation ago, cell growth and status consistent with P1 generations, with spindle cells cultured gradually reduce increased polygon cells, cell volume continues to become larger, trypsin digestion started to become difficult, the first P10 era, but also adherent cells, but basically stopped growing,trypsin digestion of about 30 min, to make cell shedding. Cell growth was not the law, can not be determined from the form. Primary cell culture Nestin staining seen a few cells, after passage only see a very small amount of positive staining cells. Umbilical cord MSCs were nestin-m RNA expression, the expression level in a significant decrease after passage. And nestin protein changes in the neural precursor cells differentiate into neural cells is consistent.Conclusion Umbilical cord mesenchymal stem cells showed positive expression of nestin-m RNA, the expression level after the passage of a significant decrease. But that does not mean reducing the amount of cells expressing the number of neural precursor cells decreased, first of all, the change in the characteristics and nestin protein changes in the neural precursor cells differentiate into neural cells is consistent; Second, this change is also likely to and horizontal cell differentiation related to this unpredictable differentiation trends are likely to affect the expression of nestin-m RNA. Therefore, to reduce the expression of nestin-m RNA may be single or combined action of two factors appeal. Gene sequences used in this study for the nestin-m RNA primers can identify these nstin-m RNA expression in neural precursor cells positive cells contain.