Bjerke T, Nielsen S, Helgestad J, Nielsen B W, Schiøtz P O
Department of Pediatrics, University Hospital of Aarhus, Denmark.
J Immunol Methods. 1993 Jan 4;157(1-2):49-56. doi: 10.1016/0022-1759(93)90069-j.
A simple and rapid method for isolating highly purified and functionally intact basophils from normal human blood is described. Blood from healthy volunteers was centrifuged through a two layer Percoll density gradient. The majority of the basophils were recovered between Percoll layers with densities of 1.070 and 1.080, constituting 45.6% of total leukocytes. Lymphocytes, monocytes and neutrophils were extracted from this fraction using a panel of mabs in a direct or indirect selection procedure using immunomagnetic beads (Dynabeads) coated with sheep anti-mouse IgG. These negative immunoselection procedures produced a high yield of basophils with a mean purity of 97.8% (range 93.0-99.5%) and 97.0% (range 96.2-99.0%) using the direct and indirect method, respectively. The cells isolated by this method are viable, release histamine to various stimuli in a normal manner, and appear morphologically normal in transmission electron microscopy.
本文描述了一种从正常人血液中分离高纯度且功能完整的嗜碱性粒细胞的简单快速方法。来自健康志愿者的血液通过两层Percoll密度梯度进行离心。大多数嗜碱性粒细胞在密度为1.070至1.080的Percoll层之间回收，占总白细胞的45.6%。使用一组单克隆抗体，通过直接或间接选择程序，利用包被羊抗小鼠IgG的免疫磁珠（Dynabeads）从该部分中提取淋巴细胞、单核细胞和中性粒细胞。这些阴性免疫选择程序分别使用直接法和间接法产生了高产率的嗜碱性粒细胞，平均纯度分别为97.8%（范围93.0 - 99.5%）和97.0%（范围96.2 - 99.0%）。通过该方法分离的细胞具有活力，能以正常方式对各种刺激释放组胺，并且在透射电子显微镜下形态正常。
