论文部分内容阅读
背景:许旺细胞是神经创伤修复的种子细胞,获取大量高纯度、高活性许旺细胞是研究的关键。目的:比较乳鼠和成年鼠许旺细胞的体外培养、纯化和形态学的差别,探讨简单可行的、可以获得高纯度许旺细胞的培养方法。方法:出生1～3dSD大鼠20只和成年大鼠10只(体质量150～200g),实验按细胞的来源分为新生组和成年组。双酶分步消化,二次接种差速分离纯化细胞;倒置显微镜观察细胞形态、贴壁速度;细胞计数,纯度计算;MTT法检测细胞增殖能力,绘制两组许旺细胞的增殖曲线,判定增殖速度;S-100免疫化学法鉴定细胞。结果与结论:乳鼠的许旺细胞贴壁速度快于成纤维细胞,成鼠的许旺细胞贴壁速度慢于成纤维细胞,两组许旺细胞纯度均达96%以上;MTT法检测两组许旺细胞增殖均活跃,由增殖曲线显示乳鼠许旺细胞增殖更快(P<0.05);S-100免疫化学反应均呈阳性。提示双酶分步消化,二次接种差速分离纯化细胞,可以获取纯度高、活性良好的许旺细胞;乳鼠许旺细胞的增殖、贴壁能力更强。
BACKGROUND: Schwann cells are seed cells repaired by nerve trauma. It is the key to study that a large number of Schwann cells with high purity and activity are obtained. OBJECTIVE: To compare the differences in the culture, purification and morphological characteristics of Schwann cells from neonatal rats and adult mice in vitro, and to explore a simple and feasible method for culturing Schwann cells with high purity. Methods: Twenty male SD rats aged 1 ~ 3 days and 10 adult rats (body weight 150 ~ 200g) were divided into neonatal group and adult group according to the origin of the cells. Double-enzyme digestion, secondary inoculation differential isolation and purification of cells; inverted microscope observation of cell morphology, adherent rate; cell count, purity calculation; MTT assay cell proliferation ability, draw two groups of Schwann cell proliferation curve, determine the proliferation Speed; S-100 immunochemistry to identify cells. RESULTS AND CONCLUSION: Schwann cells in neonatal rats adhered faster than fibroblasts, the adherent Schwann cells in fibroblasts were slower than those in fibroblasts. The purity of Schwann cells in both groups reached more than 96% The proliferation of Schwann cells were all active. The proliferative curve showed that the proliferation of Schwann cells was faster (P <0.05). The immunochemical reaction of S-100 was positive. Prompt double-enzyme digestion, secondary vaccination differential isolation and purification of cells, you can get high purity and good activity of Schwann cells; Schwann cells proliferation, adherent ability.