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最近研究显示,分枝样小胶质细胞在大脑中发挥各种作用。但是,原代培养小胶质细胞这一研究模型并不足以研究这种独特的细胞。本研究提供了一种新的研究方法:培养海马脑片(organotypic hippocampal slice cultures,OHSC),从成年鼠脑中分离分枝样小胶质细胞,并将该细胞移植到海马脑片中(去掉相应位置的细胞),得到小胶质细胞移植海马脑片(microglia-replenished OHSC,Mrep-OHSC)。移植后的小胶质细胞和海马组织融合,分枝延展,与鼠脑中的原有的分枝样小胶质细胞无法区分。从TNFα敲除小鼠中提取小胶质细胞,并移植到野生型小鼠脑片,与来自于特异性小胶质细胞TNFα敲除小鼠(CX3CR1cre/TNFαfl/fl)的脑片具有相似的特性。本研究还显示,移植到海马脑片中的小胶质细胞保持了其原有的(在体)功能和特性。从早老模型小鼠(ERCC1△/ko)中提取小胶质细胞并移植到野生型海马脑片中,这些小胶质细胞维持其原有表型并继续高表达Mac2。综上所述,本研究证明Mrep-OHSC是构建嵌合子脑片的独特的组织培养模型,可用于研究在体不同表型的小胶质细胞的功能。
Recent studies have shown that branched microglia play a variety of roles in the brain. However, this model of primary culture of microglia is not sufficient to study this unique cell. This study provides a new method of research: culturing the hippocampal slices (OHSC), separating the microglia from the adult rat brain and transplanting the cells into the hippocampal slices Corresponding cells) to obtain microglia-replenished OHSC (Mrep-OHSC). After transplantation, microglia and hippocampus tissue fusion, branch extension, and the mouse brain in the original branch-like microglia can not be distinguished. Microglial cells were extracted from TNFα knockout mice and transplanted into wild-type mouse brain slices with similar to brain slices from specific microglia TNFα knockout mice (CX3CR1cre / TNFαfl / fl) characteristic. The study also showed that microglia transplanted into the hippocampal slices retained their original (in vivo) function and character. Microglial cells were extracted from premature old mouse models (ERCC1Δ / ko) and transplanted into wild-type hippocampal slices, which maintained their original phenotype and continued to overexpress Mac2. In summary, the present study demonstrates that Mrep-OHSC is a unique tissue culture model for constructing chimeric brain slices that can be used to study the function of microglia in different phenotypes in vivo.