中国免疫学杂志
中國免疫學雜誌
중국면역학잡지
CHINESE JOURNAL OF IMMUNOLOGY
2015年
1期
86-89
,共4页
林炜明%戴爱玲%尹会方%杨小燕
林煒明%戴愛玲%尹會方%楊小燕
림위명%대애령%윤회방%양소연
巨噬细胞%磁珠分选%培养%功能鉴定
巨噬細胞%磁珠分選%培養%功能鑒定
거서세포%자주분선%배양%공능감정
Macrophages%Magnetic activated cell sorting%Culture%Functional identification
目的:从人外周血单个核细胞( PBMC)中分离单核细胞,诱导分化成巨噬细胞并鉴定其功能。方法:应用免疫磁珠法从人外周血单个核细胞中分选CD14+单核细胞,分离后的细胞用流式细胞仪检测其纯度,用含有10%人AB血清和10 ng/ml人M-CSF的IMDM培养基体外培养CD14+单核细胞,诱导分化成巨噬细胞,并进行形态特征和吞噬功能鉴定。结果:免疫磁珠法能从外周血中分离到高纯度的CD14+单核细胞,分选前CD14阳性率为10%,分选后CD14阳性率为85.8%。诱导培养7天后巨噬细胞的直径最大可达40~45μm,大部分细胞呈煎蛋状,能有效地吞噬淋巴瘤Raji细胞。结论:从外周血中分离了高纯度的CD14+单核细胞,诱导形成的巨噬细胞具有吞噬淋巴瘤细胞的功能。
目的:從人外週血單箇覈細胞( PBMC)中分離單覈細胞,誘導分化成巨噬細胞併鑒定其功能。方法:應用免疫磁珠法從人外週血單箇覈細胞中分選CD14+單覈細胞,分離後的細胞用流式細胞儀檢測其純度,用含有10%人AB血清和10 ng/ml人M-CSF的IMDM培養基體外培養CD14+單覈細胞,誘導分化成巨噬細胞,併進行形態特徵和吞噬功能鑒定。結果:免疫磁珠法能從外週血中分離到高純度的CD14+單覈細胞,分選前CD14暘性率為10%,分選後CD14暘性率為85.8%。誘導培養7天後巨噬細胞的直徑最大可達40~45μm,大部分細胞呈煎蛋狀,能有效地吞噬淋巴瘤Raji細胞。結論:從外週血中分離瞭高純度的CD14+單覈細胞,誘導形成的巨噬細胞具有吞噬淋巴瘤細胞的功能。
목적:종인외주혈단개핵세포( PBMC)중분리단핵세포,유도분화성거서세포병감정기공능。방법:응용면역자주법종인외주혈단개핵세포중분선CD14+단핵세포,분리후적세포용류식세포의검측기순도,용함유10%인AB혈청화10 ng/ml인M-CSF적IMDM배양기체외배양CD14+단핵세포,유도분화성거서세포,병진행형태특정화탄서공능감정。결과:면역자주법능종외주혈중분리도고순도적CD14+단핵세포,분선전CD14양성솔위10%,분선후CD14양성솔위85.8%。유도배양7천후거서세포적직경최대가체40~45μm,대부분세포정전단상,능유효지탄서림파류Raji세포。결론:종외주혈중분리료고순도적CD14+단핵세포,유도형성적거서세포구유탄서림파류세포적공능。
Objective:To isolate monocytes from human peripheral blood mononuclear cells( PBMC) ,induce macrophages,and identify the function of macrophages.Methods:Monocytes were isolated from PBMC using magnetic activated cell sorting( MACS) anti-CD14 microbead.Sorted CD14+and CD14-cells were checked by flow cytometer to evaluate the efficiency of sorting.The sorted CD14+cells were cultured in IMDM media with 10%human AB serum and 10 ng/ml M-CSF for 7 days to generate macrophages,which were identified by morphological features and phagocytosis function.Results:A high purity of monocytes was obtained by MACS anti-CD14 microbead.The percentage of CD14+cells was 10% and 85.8% before and after sorting, respectively.The macrophages were approximately 40-45 μm in maximum diameter and had the fried egg colony morphological features after 7 days culture.The lymphoma ( Raji) cells were efficiently engulfed by macrophages.Conclusion: The high purity of CD14+monocytes is isolated from PBMC and monocyte-derived macrophages efficiently engulfed lymphoma cells.