人胎盘间充质干细胞对脐血淋巴细胞转化的影响  

Effect of human placenta derived mesenchymal stem cells on cord blood lymphocyte transformation

作　　者：李长东 [1] 张为远 [2] 李荷莲 [3] 江小霞 [4] 张毅 [4] 毛宁 [4] 

机构地区：[1]100026,首都医科大学附属北京妇产医院妇科 [2]100026,首都医科大学附属北京妇产医院产科 [3]吉林大学第二医院妇产科 [4]军事医学科学院基础医学研究所细胞生物研究室

出　　处：《中华医学杂志》2005年第24期1704-1707,共4页National Medical Journal of China

摘　　要：目的从人胎盘中分离间充质干细胞(MSC),研究胎盘MSC在脐血移植中的免疫调节作用。方法分离人胎盘组织灌流液中单个核细胞(MNC)贴壁培养、集落筛选,鉴定表面标志及纯度,向脂肪、成骨诱导鉴定多向分化潜能。应用3HTdR掺入法,观察胎盘MSC对成人外周血淋巴细胞及脐血淋巴细胞的免疫抑制作用。将不同数量胎盘MSC分别与成人外周血和脐血淋巴细胞共培养,加入植物血凝素(PHA)刺激淋巴细胞增殖,观察胎盘MSC对淋巴细胞转化的影响。结果从人胎盘组织中分离获得的贴壁细胞,具有同骨髓MSC相同的细胞表型及多向分化能力,可以显著抑制成人外周血和脐血淋巴细胞转化,抑制作用与MSC的数量呈正相关。与无MSC作用相比,当MSC为2×105/孔时对成人外周血和脐血淋巴细胞转化的抑制率分别为79.97%和64.06%,而当MSC为1.25×104/孔时抑制率则分别为46.83%和12.19%。结论从人胎盘中可以成功地分离出MSC,利用其对脐血淋巴细胞的免疫调节作用,除了可以作为与脐血造血干/祖细胞相适应的培养基质,还有望降低脐血移植中困扰已久的移植物抗宿主疾病(GVHD)问题,促进脐血移植的广泛应用。Objective To study the effect of human placenta derived mesenchymal stem cells (MSCs) on the immune function of lymphocytes derived from human umbilical cord blood. Methods Mononucleated cells (MNC) were isolated from human placenta tissue perfusate by density gradient fractionation. Individual colonies were selected and cultured. The culture-expanded cells were characterized by immune phenotyping so as to identify the MSCs. The MSCs were cultured under conditions promoting differentiation to osteoblasts or adipocytes. MNCs were isolated from adult peripheral blood and human umbilical cord blood and cultured, then the adherent cells were excluded and the suspended cells, lymphocytes, were inoculated in the culture fluids of MSCs of different concentrations and phytohemagglutinin (PHA), a nonspecific mitogenic stimulant, was added for 84 hours (MSC+PHA groups), then 3H-thymidine deoxyribose (3H-TdR) was added for 12 hours. The cells were collected and scintillation counter was used to calculate the counts per minute (cpm). Pure lymphocytes without MSC and stimulated by PHA were used as control group (non-MSC Group) and pure lymphocytes and pure MSCs without PHA were used as blank control groups (non-PHA Group). Results From human placenta MSCs were successfully isolated and exhibited fibroblast-like morphology. Flow cytometric analysis showed that the placental MSCs were a homogeneous cell population devoid of hematopoietic cells positive for CD29, CD44, CD73, CD105, CD166, and HLA-ABC positive and negative for CD34, CD45, and HLA-DR. They could be induced into adipocytes or osteocytes. The cpm value of the non-MSC Group was 171 855±31 454, significantly higher than that of non-PHA Group (26 453±5268). The cpm values of the different concentrations MSC+PHA groups were all significantly lower than that of non-MSC Group in a dose-dependent manner; when the dose of MSCs was 2 × 105 the suppression rate was 79.97% in PB and 64.06% in UCB. Conclusion MSCs derived from postpartum human placenta, an important and nove

关 键 词：淋巴细胞转化 间充质干细胞 脐血造血干/祖细胞 人外周血淋巴细胞 移植物抗宿主疾病 免疫调节作用 脐血淋巴细胞 成人外周血 人胎盘组织 脐血移植 多向分化潜能 TdR掺入法 免疫抑制作用 淋巴细胞增殖 MSC 单个核细胞 细胞共培养 

分 类 号：R392[医药卫生—免疫学]