Szabó C A, Deli M A, Ngo T K, Joó F
Laboratory of Molecular Neurobiology, Hungarian Academy of Sciences, Szeged, Hungary.
Neurobiology (Bp). 1997;5(1):1-16.
To study the blood-brain barrier in vitro pure cerebral endothelial cell cultures, without contaminating cells have to be obtained. Most other cell types besides endothelial cells can be pericytes, a few astrocytes, some smooth muscle cells, fibroblasts and meningeal cells. Careful removal of large vessels and meninges during the dissection and the optimal duration of enzymic digestions can reduce the ratio of contaminant cells. In order to further increase the purity of the culture endothelial cells can be subcloned, however, this is not useful for cells of every species. An alternative choice in cultures from rat is to perform a selective cytolysis by complement and monoclonal anti-Thy 1.1 antibody to eliminate pericytes and astrocytes. The presence of growth factors and the type of serum are also important for successful endothelial cell cultures. With the combination of the cytolysis of contaminating cells and the use of plasma-derived serum, the culturing of pure primary cerebral endothelial cells was successful.
为了在体外纯脑内皮细胞培养物中研究血脑屏障，必须获得无污染物的细胞。除内皮细胞外，大多数其他细胞类型可能是周细胞、一些星形胶质细胞、一些平滑肌细胞、成纤维细胞和脑膜细胞。在解剖过程中小心去除大血管和脑膜以及酶消化的最佳持续时间可以降低污染物细胞的比例。为了进一步提高培养物的纯度，内皮细胞可以进行亚克隆，然而，这对每个物种的细胞并不都有用。在大鼠培养物中的另一种选择是通过补体和单克隆抗Thy 1.1抗体进行选择性细胞溶解，以消除周细胞和星形胶质细胞。生长因子的存在和血清类型对内皮细胞培养的成功也很重要。通过结合污染物细胞的细胞溶解和使用血浆衍生血清，成功培养了纯原代脑内皮细胞。
