| Objective: To explore the neural stem cells and temozolomide for livingconditions of cerebral glioma in rats. Methods: The experiment is divided intothree parts. The first part: the isolation, culture and identification of rat neuralstem cells.(1) selection of12~14days gestational age of fetal rats, isolatedfrom these fetal mouse Neural stem cells (NSCs), then using serum-free culturedevelop and extend the way;(2) to isolate the Nestin identification of neuralstem cells by immunofluorescence, and purification of3~4generation ofneural stem cells after heavy ball, and training, make its active is the best state,and adjust the cell density of4x106/20ul. The second part: rat C6gliomamodel.(1) take cryopreserved C6glioma cells for recovery, training,identification of cell survival, take the logarithmic phase cells, at the same time,adjust the cell density is106/20ul for use, and will be part of unused C6gliomacells cryopreserved;(2) The exponential phase of C6glioma cells, using brainstereotactic apparatus to rat caudate nucleus vaccination the C6glioma cells,build90C6rat glioma model, to observe the postoperative survival quality andsurvival time of rats, and record, there are8only accidental death in rats. Thethird part: the neural stem cells and temozolomide impact on the survivalcondition of glioma rats. Neural stem cells and temozolomide treatment of ratC6glioma,7days after the second part, the remaining82model rats randomly selected two to death, HE staining in tissue of profile control cranial tumorremoved. The remaining80were randomly divided into four groups: controlgroup (group A), neural stem cells into group (group B), temozolomide group(group C), temozolomide associated neural stem cell group (group D); Group A:anesthesia and fixed, in stereotaxic coordinates to the first frame guided salineinjections into tumor within20ul. Group B: anesthesia fixed, with head ofstereotaxic frame under the guidance of a total of4x106within20ul neuralstem cells injected into the tumor. Group C: using the lavage needle to rats forpumping temozolomide:50mg/kg, take5days. Group D: anesthesia fixed,under the guidance of stereotaxic frame head will be a total of4x106within20ulneural stem cells injected into the tumor, neural stem cells injected into the nextday to start to rats with gastric needle injection temozolomide50mg/kg, take5days. Quality of life and survival of rat were observed and recorded, survival ofrat survival analysis, and map ICONS. Death in rat models of profile controlafter cranial probe and tumor removed, washed with such as infiltration ofphysiological saline, to look at the four groups of tumor, remove the tumor withelectronic weighing scales, and records, statistical analysis, and map ICONS.After weighing the tumor tissue for HE staining. Results: The first part, cultureand identification of rat neural stem cells: the separation of fetal rat neural stemcells, a large number of cells in the growth of the single cell suspension stateand vary in size and form, cells of different amount of balls;Immunofluorescence staining Nestin positive, and confirmed the cells for neural stem cells with active. The second part，rat C6glioma model isestablished in this paper:(1) the C6glioma cells in logarithmic phase, the cellgrowth closely packed, the density is evener, stick a wall is fast; Most of thepolygon, spindle.(2) build90SD male rat C6glioma model: intraoperativedied in the accident,2rats in operation on the third day4rats died ofintracranial hematoma,2rats with only5days unexplained death; The rest ofthe82model rats into experimental processing and analysis. Modeling rats2days after the operation, the feed itself, but to eat and drink less preoperative,fur is not neat, color luster, depression;6days after inoculation, aggression,escape behavior are enhanced. The third part， neural stem cells andtemozolomide effect on the survival condition of glioma rats: neural stem cellsinto group (group B), temozolomide group (group C), and combined neuralstem cells temozolomide group (group D) compared with control group (groupA) quality of life improved; A, B, C, D four groups of average lifetime are:23.700days,28.600days,28.900days,29.500days. Kaplan Meier-betweenthe two method analysis: B, C, D stage was extended, the three groups werecompared with group A survival statistics difference (p <0.05), but the B, C, Dbut survival is no statistical difference between three groups (p>0.05). Eachprofile control after the death of rat cranial observed in each group are in theright hemisphere slightly front ovalisation tumor growth. A, B, C, D fourgroups of tumor weight, respectively is:104.105±9.877mg，95.175±9.387mg，95.870±9.542mg，95.905±8.807mg; Line between groups of analysis of variance,F=4.061,P=0.010,the difference was statistically significant,can think of all the weight of the four groups is not the same; T test arecompared, and two is adopted by the t test to compare two hints: the tumorweight of the rats of group A is B, C, D three groups are heavier, morestatistically significant (p <0.05); B, C, D group of pairwise comparisonbetween tip: tumor weight is no statistical difference (p>0.05). Four groups ofglioma tissues HE staining slices have see the growth of tumor cells isextremely active, accompanied by a certain invasive; Normal brain tissue andtumor border to see a large number of infiltrating growth of tumor cells, there isa clear boundary, increasing enlargement of tumors within the blood vessels.Conclusions:(1)Identified by neural stem cells can use Nestin, afteridentification of neural stem cells can be used in the experiment.(2)Stereotactic method was adopted to construct the SD rat C6glioma model,stability, high reliability, the model is an ideal animal model of cerebral glioma,suitable for the experimental research for the treatment of gliomas.(3)Neural stem cells and temozolomide can prolong the survival of glioma rats,improve the quality of survival, weight the tumor.(4)Neural stem cells joint temozolomide is relatively simple for mo thiazoleamine or neural stem cells to treat glioma rats, survival, quality of life, and thetumor weight has no obvious improvement. |
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