CAJ | 학술논문

目的探讨稳定的成人视乳头星形胶质细胞原代纯化培养方法及其理想细胞模型.方法实验研究.解剖显微镜下分离出人眼视乳头筛板组织,将其剪切成4～6个小组织块,放置于含有DMEM/F12培养基的培养皿中培养.8～ 10周后去除组织块,以0.25％胰蛋白酶消化细胞,改用星形胶质细胞选择性培养基,经两次传代后,通过细胞形态学和胶质细胞原纤维酸性蛋白、神经细胞黏附分子免疫荧光染色,观察纯化培养的星形胶质细胞生长情况并进行细胞鉴定.结果组织块接种2～3周后,有细胞相继从其边缘爬出,迅速向周围分裂生长,其移行过程明显.胰酶消化后细胞的形态多呈扁平星形或多角形,也可见长梭形细胞.改用星形胶质细胞选择性培养基培养并传代后,细胞几乎均呈扁平星形,并表达胶质细胞原纤维酸性蛋白和神经细胞黏附分子.结论精确分离筛板组织并将小组织块定位于培养液边缘(培养液并未完全覆盖皿底),有利于组织块贴壁生长,是获取人原代星形胶质细胞的前提条件；星形胶质细胞选择性培养基是获得原代纯化星形胶质细胞的简便有效方法.
목적 탐토은정적성인시유두성형효질세포원대순화배양방법급기이상세포모형.방법 실험연구.해부현미경하분리출인안시유두사판조직,장기전절성4～6개소조직괴,방치우함유DMEM/F12배양기적배양명중배양.8～ 10주후거제조직괴,이0.25％이단백매소화세포,개용성형효질세포선택성배양기,경량차전대후,통과세포형태학화효질세포원섬유산성단백、신경세포점부분자면역형광염색,관찰순화배양적성형효질세포생장정황병진행세포감정.결과 조직괴접충2～3주후,유세포상계종기변연파출,신속향주위분렬생장,기이행과정명현.이매소화후세포적형태다정편평성형혹다각형,야가견장사형세포.개용성형효질세포선택성배양기배양병전대후,세포궤호균정편평성형,병표체효질세포원섬유산성단백화신경세포점부분자.결론 정학분리사판조직병장소조직괴정위우배양액변연(배양액병미완전복개명저),유리우조직괴첩벽생장,시획취인원대성형효질세포적전제조건；성형효질세포선택성배양기시획득원대순화성형효질세포적간편유효방법.
Objective To establish a method of purifying and characterizing adult astrocytes from optic nerve head ( ONH ).Methods Experimental study.The lamina eribiosa tissue from ONH of human eye was isolated under anatonic microscopy,and then 4 to 6 little explants were incubated in each culture plate containing culture medium DMEM/F12.After 8 to 10 weeks,the cells were renoved by digesting cells with 0.25％ trypsogen. Selective astrocyte culture medium is subsequently used. After two passages,astrocytes were identified by the observation of cell morphology and immunofluorescent staining of GFAP and NCAM.Results After 2 to 3 weeks of explants planting,cells showed an obvious migration procession by crawling in succession from the verge of the explants and rapidly splitting.Most cells displayed a flat star shape or polygon after digested with trypsogen.Several cells are long fusiformis.Almost all cells presented a flat star shape and simultaneously expressed GFAP and NCAM when the cells cultured with selective astrocyte culture medium.Condusions Cultured human ONH astrocytes can be obtained by precisely separating lamina cribiosa and placing the explants on the margin of culture medium,a method that promotes cell adherence.Using selective astrocyte culture medium is very effective and convenient in purifying primary astrocytes.