Wysocki L J, Sato V L
Proc Natl Acad Sci U S A. 1978 Jun;75(6):2844-8. doi: 10.1073/pnas.75.6.2844.
We have developed a simple method for the fractionation of T and B lymphocytes. Plastic dishes coated with antibodies specific for mouse Ig selectively bind splenic B lymphocytes. The adherent cells are easily removed by gentle pipetting; both adherent and nonadherent populations retain immunologic function. In a typical experiment, when 3 X 10(7) splenic lymphocytes were added to a 100 X 15 mm plastic dish coated with microgram quantities of anti-Ig, 98 % of the nonadherent cells were Ig negative and 97% of the adherent cells were Ig positive. The method is sufficiently sensitive to allow detection and separation of cell types comprising as little as 2% of the total population and can be modified to allow the selection of cells by a double-antibody procedure. We believe that the plastic dish method will be generally useful for fractionating cells on the basis of their cell surface antigens.
我们已经开发出一种简单的方法来分离T淋巴细胞和B淋巴细胞。用特异性针对小鼠Ig的抗体包被的塑料培养皿可选择性地结合脾脏B淋巴细胞。通过轻柔吹打很容易去除贴壁细胞;贴壁细胞群和非贴壁细胞群都保留免疫功能。在一个典型实验中,当将3×10⁷个脾脏淋巴细胞加入到一个用微克量抗Ig包被的100×15mm塑料培养皿中时,98%的非贴壁细胞Ig呈阴性,97%的贴壁细胞Ig呈阳性。该方法灵敏度足够高,能够检测和分离占总细胞群低至2%的细胞类型,并且可以进行改进以通过双抗体程序选择细胞。我们认为,基于细胞表面抗原分离细胞时,塑料培养皿法将具有广泛的用途。