Davidson W F, Parish C R
J Immunol Methods. 1975 Jun;7(2-3):291-300. doi: 10.1016/0022-1759(75)90026-5.
A procedure is described for simultaneously removing red cells and dead cells from lymphoid cell suspensions, based on the observation that when populations of lymphoid cells are centrifuged on a mixture of Isopaque/Ficoll, dead cells and red cells sediment whereas viable cells float. The technique very efficiently removed red cells from a wide range of lymphoid cell suspensions and eliminated lymphocytes killed by mechanical stress, by antibody and complement and by prolonged tissue culture. The depletion of red cells was greater than 99% and the recovery of viable lymphocytes usually greater than 90%, the resulting cell suspensions being around 95-100% viable. The immunological activity of B cells, helper T cells and cytotoxic T cells virtually unimpaired by the separation procedure.
本文描述了一种从淋巴细胞悬液中同时去除红细胞和死细胞的方法,该方法基于以下观察结果:当淋巴细胞群体在异泛影葡胺/聚蔗糖混合物上离心时,死细胞和红细胞会沉淀,而活细胞会漂浮。该技术非常有效地从多种淋巴细胞悬液中去除了红细胞,并消除了因机械应力、抗体和补体以及长时间组织培养而死亡的淋巴细胞。红细胞的去除率大于99%,活淋巴细胞的回收率通常大于90%,所得细胞悬液的活力约为95%-100%。B细胞、辅助性T细胞和细胞毒性T细胞的免疫活性在分离过程中几乎未受影响。