CAJ | 학술논문

目的：建立一种简易高效的大鼠脊髓小胶质细胞原代培养方法。方法：取新生SD大鼠的脊髓进行脊髓胶质细胞的原代混合培养，第3天换液一次，此后一直不换液。至第10天左右脊髓小胶质细胞长满后，在恒温摇床上振摇（37℃、180 rpm、1~2 h），并采用差速粘附20~40 min，纯化脊髓小胶质细胞。用Iba1和CD11b的免疫荧光染色对分离纯化24 h后的小胶质细胞进行纯度鉴定。结果：成功分离纯化大鼠脊髓小胶质细胞。细胞体呈圆形或者梭形，折光性很强。Iba1和CD11b免疫荧光鉴定结果显示小胶质细胞的纯度≥95%。结论：采取营养剥夺，以及振摇和差速粘附法建立了一种高产量、高纯度的脊髓小胶质细胞培养方法。
목적：건립일충간역고효적대서척수소효질세포원대배양방법。방법：취신생SD대서적척수진행척수효질세포적원대혼합배양，제3천환액일차，차후일직불환액。지제10천좌우척수소효질세포장만후，재항온요상상진요（37℃、180 rpm、1~2 h），병채용차속점부20~40 min，순화척수소효질세포。용Iba1화CD11b적면역형광염색대분리순화24 h후적소효질세포진행순도감정。결과：성공분리순화대서척수소효질세포。세포체정원형혹자사형，절광성흔강。Iba1화CD11b면역형광감정결과현시소효질세포적순도≥95%。결론：채취영양박탈，이급진요화차속점부법건립료일충고산량、고순도적척수소효질세포배양방법。
Objective: To establish a simple and highly efficient primary culture method for microglia from rat spinal cord. Methods: Mixed glial cells were obtained from the spinal cord of newborn SD rats. On day 3, the culture medium was changed until the cultured microgila became fused on day 10. The microglia were purified using shocking methods (37 ℃, 180 rpm, 1~2 h) with different adhesion time (20~40 min). The isolated and purified microglia were identified with Iba1 and CD11b after 24 h. Results: The rat spinal cord microglia were successfully isolated and purified. The cultured microglia presented round or fusiform, highly refractive morpholo-gy. Iba1 and CD11b fluorescence tests showed that the purity of microglia was more than 95%. Conclusion:Using nutrition deprivation, Shock and different adhesion time methods, a primary spinal cord microglia culture method with high yield and purity was established, providing a useful tool for studying rat spinal cord microglia in vitro.