| Objective to study the Content and distribution characteristics of CD133-positiveglioma stem cells (GSC) in human glioma-bearing of nude mice with flowcytometry andimmunohistochemistry,and to explore the relationship between the content of GSC andtumor size,inorder to lay the basis for the subsequent work of molecular imaging of gliomastem cells of MR.Material and methods30BALB/c nude mice with age4-5weeks, weight of15to20g, glioma stem cell lines of SU2cell inoculated to nude mice subcutaneously afterculture,then observed the tumors. According to the tumor diameter,they were divided intotwo groups,one was group5mm and the other was group10mm,animals were sacrificedafter MRI (T1WI,T2WI and enhanced T1WI) scanning.then removed the tumors, anddivided into two equal parts, one for flow cytometric to observed the content of CD133-positive GSC of tuomors, one for immunohistochemistry to analyse the content anddistribution of CD133and nestin-positive GSC of tumors, and the relationship betweentumor size,in addition,CD34Marked Weidner method to measure the MVD to analyse therelationship between the content of the CD133-positive GSC and MVD. SPSS16.0_statistical software package for statistical analysis, results were expressed as mean x±_standard deviation (x±s), the content of CD133and nestin-positive cell and MVD wereexplored respectively with t text, the correlation of CD133positive cells and MVD withlinearcorrelation test,P<0.05considered statistically significant difference.Result:(1) inoculation SU2cells into30nude mice of armpit,four of them was deathdue to anesthesia accident,the remaining26were tumorigenic.6were removed becauselongest diameter of tumor were greater than10mm,the remaining20nude mice weredivided into two groups,group5mm and group10mm according to the tumor diameter. thecontent of CD133and nestin was reduced with the increase of the diameter, there were statistical significant of the two groups (t=3.23,P=0.005), consistent with the Flowcytometry.(2) cancer stem cells were mostly nested, around the blood vessels and necrosisthe number of stem cells in tumor marginal zone were more than center area.(3)The largerthe diameter of the tumors the lower of MVD levels,there were statistical significant of thetwo groups (t=3.16,p=0.005).there was no correlation between the content of CD133positive cell and MVD.neovascular was more likely concentrated around the tumor marginand necrosis.(2) MRI showed that the tumor was oval, the signal of T1WI was similar tothe muscle,T2WI showed high signal, both groups were enhanced, group10mm can seeno enhancement area of necrosis of tumor.(3)the content of CD133and nestin-positiveGSC were reduced with the increase of the tumor (t=3.23, P=0.005).(4) flow cytometryshowed the content of CD133-positive GSC of group5mm was5.03±2.53(%) and group10mm was2.42±0.79(%),Immunohistochemistry showed the content of CD133-positivecell of group5mm was4.02±1.15(%) and group10mm was2.74±0.51(%), the contentof CD133-positive cell with two methods showed no difference (t=1.15,P=0.27, t=1.14,P=0.27).(5) the CD133-positive GSC was located in the margin of tumor,and around thelarger blood vessels,nested and gathered piles.The smaller the diameter of the tumors,themore content of GSC. more phenomenon of niche of stem cell.GSC distributed less inCancer Center Regional.(6) The larger the tumor, the less content of MVD(t=3.16,p=0.005). No significant correlation between the content of CD133-positive cell and MVD(r=0.562,P=0.091).Conclusion:(1) the formation and stability of tumor was high of SU2cell line ininoculation axillary subcutaneous of nude mice,which can be used for the CD133positiveGSC molecular imaging of animal models in MR;(2) the content of CD133-positive GSCwas reduced with the increase of tumor;(3) CD133-positive GSC distributed around theblood vessels of the tumor edge.(4)the content of CD133-positive cell and MVD was nosignificant correlation. |
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