Isolation, Cultivation And Identification Of Umbilical Cord Blood Stem Cells In Vitro And Efficacy Analysis Of Usingfor Cerebral Hemorrhage Sequel

The study included the following two parts:Part Ⅰ The isolation, culture and identification of Cord blood stem cells in vitro.Objective umbilical cord blood stem cells were isolated by adopting the densitygradient centrifugation combining with direct stick wall, and gave the original generation andsubculture in vitro, and observed cells form, identified the ability of proliferation and whetherexpressed neural stem cells specific antigen. Methods according to the method of document[1], obtained umbilical cord blood stem cells by using stem cells liquid, after originalgeneration and subculturing by using the DMEM/F12medium in vitro, observed cells form,identified the ability of proliferation and whether expressed neural stem cells specific antigenby immune cells fluorescent chemical dyeing methods. Results Primary culture of theumbilical cord blood stem cell, cloning ball contained positive cells that expressed Nestin andNSE, although the umbilical cord blood stem cells handed from generation to generationseveral times, Umbilical cord blood stem cells could proliferate stably, proportion of thepositive cells that expressed Nestin antigen in cloning ball had no obvious variation.Conclusion The experiment proved that the neural stem cells could be separate andcultivated from umbilical cord blood stem cells, after repeatedly passaging some cellsexpressed NSE, Thus, lay the solid foundation for the clinical treatment of nervous systemdiseases.Part Ⅱ The analysis of curative effect about umbilical cord blood stem cells transplantation in the treatment of cerebral hemorrhage sequel.Objective From the angle of brain cells regeneration, by both at home and abroad about thebasis of the research on the theory of the induction, differentiate into neural stem cells aboutumbilical cord blood stem cells,migration and regeneration of neural stem cells aboutmammals in vitro, This subject mainly studies transplanting umbilical cord blood stem cellsinto sequel patients of cerebral hemorrhage，through recovery of the neural function and thealteration of Cell growth factor of cerebrospinal fluid of the treatment group, to evaluateobjectively the curative effect of Cerebral hemorrhage sequel, so as to explore a new way tothe treatment of intracerebral hemorrhage. Methods60people were randomly divided intotwo groups: The treatment group: cord blood stem cells isolated were poured into spinal canalthrough the lumbar puncture, and gave appropriate dehydration and neural nutrition medicineto treatment. The control group: in addition to not to give stem cell transplantation, treatmentwith more than the treatment group. Two groups, limb function was scored3,6,9monthsafter treatment using Scandinavia Stroke Scale rating; detected Cell growth factor ofcerebrospinal fluid before treatment and1,2,3weeks after treatment. Results1. Twogroups, limb function was scored3,6,9months after treatment using Scandinavia StrokeScale rating, Results show that the treatment group was significantly higher than the controlgroup. P <0.05difference had statistical significance.2. Stem cell transplantation group,Cerebrospinal fluid Cell growth factor increased significantly after treatment, P <0.05difference had statistical significance. Conclusion umbilical cord blood stem celltransplantation in the treatment of cerebral hemorrhage sequel had obtained the certaincurative effect, gave the treatment of diseases of the nervous system bring new dawn.