PURIFICATION OF HUMAN BLOOD BASOPHILS BY NEGATIVE SELECTION USING IMMUNOMAGNETIC BEADS

被引：39

作者：

BJERKE, T ^{[1
]}

NIELSEN, S ^{[1
]}

HELGESTAD, J ^{[1
]}

NIELSEN, BW ^{[1
]}

SCHIOTZ, PO ^{[1
]}

机构：

[1] AARHUS UNIV,DEPT CELL BIOL,DK-8000 AARHUS,DENMARK

来源：

JOURNAL OF IMMUNOLOGICAL METHODS | 1993年 / 157卷 / 1-2期

关键词：

BASOPHIL; HUMAN; NEGATIVE SELECTION; IMMUNOMAGNETIC BEAD; HISTAMINE RELEASE;

D O I：

10.1016/0022-1759(93)90069-J

中图分类号：

Q5 [生物化学];

学科分类号：

071010 ; 081704 ;

摘要：

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.

引用

页码：49 / 56

页数：8