【摘要】 提高肿瘤浸润淋巴细胞（ＴＩＬ）的增殖能力和杀伤活性，是其临床应用前需解决的关键问题。为此，作者对ＴＩＬ的分离和培养条件进行了探讨，建立了以机械分散法，０．０５％胶原酶和０．００３％ＤＮＡ酶室温消化６小时加冷消化１８小时的酶消化法和７５％与１００％Ｆｉｃｏｌｌ非连续密度梯度离心法相结合的分离纯化ＴＩＬ方法。分离出的ＴＩＬ在体外条件培养基中多数能扩增到１０９以上的应用标准，ＮＫ、ＬＡＫ活性分别可达５８．３±１１．７％和４８．６±１０．６％，其中发挥主要作用的是ＣＤ８Ｔ细胞。因此，本法不失为一种有效的分离培养ＴＩＬ的方法。它的建立为应用ＴＩＬ治疗恶性肿瘤奠定了良好的实验基础。更多还原

【Abstract】 The key to preparing TILs for clinicalapplication is to enhance its capacity of proliferationand cvtotoxicity,For this purpose we have rnade astudy of methodology and established an effective wayconsisting of 3 processes to separate and culture TILs.It included:(1) mechanical splitting; (2) digestingwith 0. 05%collagenase Ⅱ and 0. 003%DNAase Ⅰmixture in room temperature for 6 hrs and 18hrs incool , and(3) centrifuging with 75%and 100%Ficollnon-continuous grade density,The separated TILswere suspended in conditional culture medium for pro-liferation. Most of them reached the amount of 109 atthe end of culture,which was essential for clinicaltherapy. Their NK and LAKactivities were 56.3±11.7%and 48.6%±10.6%respectively,in which theCD5 T cells played an important role,The resultssuggest this a perfect way for separating and culturingTILs.更多还原