Orlandi M, Bartolini G, Chiricolo M, Minghetti L, Franceschi C, Tomasi V
Prostaglandins Leukot Med. 1985 May;18(2):205-16. doi: 10.1016/0262-1746(85)90020-4.
We have developed a technique to isolate monocytes from human peripheral blood. The technique takes special care of completely eliminating platelets which are usually present in other preparations. Monocytes obtained in good yields (2.5-5.0 X 10(5) cells/ml blood), were found to be 70-80% pure on the basis of morphological and histochemical criteria. Contamination was largely due to the presence of lymphocytes. Monocytes were incubated in the presence or absence of arachidonic acid and TXB2, PGE2 and 6-keto-PGF1 alpha were measured by specific and sensitive radio-immunoassays. It was found that when cells were incubated for up to 1 hr, the production of PGs was low or absent even in the presence of 10 microM arachidonic acid in the incubation medium. However, when incubations were carried out for 24 hrs in the presence of at least 1% fetal calf serum a dramatic increase in TXB2 production occurred, with levels as high as 150 ng X 10(6) cells. The ratio TXB2/PGE2 was around 3, while 6-keto-PGF1 alpha was produced at a much lower level. In the same conditions, when care was taken to evaluate PGs already present in fetal serum and/or cross reactivity due to media generally employed, purified human lymphocytes appeared unable to produce detectable levels of the three PGs tested.
我们已经开发出一种从人外周血中分离单核细胞的技术。该技术特别注意完全去除通常存在于其他制剂中的血小板。获得的单核细胞产量良好（2.5 - 5.0×10⁵个细胞/毫升血液），根据形态学和组织化学标准，发现其纯度为70 - 80%。污染主要是由于淋巴细胞的存在。将单核细胞在有或没有花生四烯酸的情况下孵育，通过特异性和灵敏的放射免疫测定法测量TXB₂、PGE₂和6 - 酮 - PGF₁α。发现当细胞孵育长达1小时时，即使在孵育培养基中存在10微摩尔花生四烯酸，PG的产生也很低或没有。然而，当在至少1%胎牛血清存在下进行24小时孵育时，TXB₂的产生显著增加，水平高达150纳克/10⁶个细胞。TXB₂/PGE₂的比值约为3，而6 - 酮 - PGF₁α的产生水平要低得多。在相同条件下，当注意评估胎牛血清中已经存在的PG和/或由于通常使用的培养基导致的交叉反应性时，纯化的人淋巴细胞似乎无法产生可检测水平的三种测试PG。
