Chihara J, Kurachi D, Yamamoto T, Yamada H, Wada T, Yasukawa A, Nakajima S
Fourth Department of Internal Medicine, Kinki University School of Medicine, Osaka, Japan.
Allergy. 1995 Jan;50(1):11-4. doi: 10.1111/j.1398-9995.1995.tb02477.x.
Eosinophils were isolated by the three methods of CD16-negative depletion: 1) magnetic beads, 2) fluorescence-activated cell sorter (FACS), and 3) complement reaction. Their purity, yield, and viability were compared. The second procedure produced well purity and viability (94.65 +/- 1.51% and 94.98 +/- 1.40%, respectively) but low yield of eosinophils (65.47 +/- 2.47%). The viability of cells obtained by the third procedure was not efficient (80.83 +/- 2.85%), while the purity and the yield were efficient (96.23 +/- 1.09% and 90.75 +/- 1.72%, respectively). In conclusion, the magnetic beads method (purity: 98.02 +/- 0.45%, yield: 91.05 +/- 2.43%, viability: 97.57 +/- 0.37%) was the most advantageous of these three procedures. Moreover, in the functional assay, radical oxygen products from eosinophils isolated by the procedure with complement reaction were less than with the magnetic beads or FACS procedures.
通过三种CD16阴性去除方法分离嗜酸性粒细胞:1)磁珠法,2)荧光激活细胞分选仪(FACS)法,3)补体反应法。比较了它们的纯度、产量和活力。第二种方法产生的嗜酸性粒细胞纯度和活力良好(分别为94.65±1.51%和94.98±1.40%),但产量较低(65.47±2.47%)。通过第三种方法获得的细胞活力不佳(80.83±2.85%),而纯度和产量较好(分别为96.23±1.09%和90.75±1.72%)。总之,磁珠法(纯度:98.02±0.45%,产量:91.05±2.43%,活力:97.57±0.37%)是这三种方法中最具优势的。此外,在功能测定中,通过补体反应法分离的嗜酸性粒细胞产生的活性氧产物比磁珠法或FACS法少。