人外周血及脐血树突状细胞的体外分离培养

Isolation and Inculation of Dendritic Cells from Human Peripheral Blood or umbilical Cord Blood

取正常人外周血或脐血，经淋巴细胞分离液分离，取中间白膜层，培养板中进行粘附，粘附细胞加培养液和细胞因子（外周血加GM－CSF和IL－4，脐血加GM－CSF和TNF－α）培养，对其形态、表型和功能分别进行鉴定和测定。结果表明约经过1周左右培养，悬浮细胞表现为典型的DC形态，带有毛刺样凸起，经DC单克隆抗体染色后用流式细胞仪测定脐血72％为DC，外周血93％为DC，并且可以刺激同种异体淋巴细胞的增殖反应。所以通过这样的不同细胞因子组合可以从人外周血和脐血中诱导培养出大量的DC细胞，为其进一步的研究奠定了基础。

Normal human peripheral blood or neonatal cord blood were isolated by ficoll centrifijgation andthe interface cells were harvested and cultured in plates.The adherent cells were then cultured withdifferent cytokine recombinantion(peripheral blood with GM-CSF+IL-4;cord blood with GMCSF+TNF-a). After one week culturing,the cells were checked by morphological.FACS and function assay. Most suspended cells exhibite distinctive morphological fearres of DC and they can alsostiumlate proliferation of allogeneic lymphcytes. These results indicate that by the different cytokinescombinantion,large numbers of DCs can be obtained from human peripheral blood or cord blood.