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Comparative research of dendritic cells cultured from mice bone marrow with different ways
Quan-Chu Wang, Zhi-Hua Feng, Yong-Xing Zhou, Qing-He Nie, Chu-Qiou Hao, Jiou-Ping Wang
Quan-Chu Wang, Zhi-Hua Feng, Qing-He Nie, Chu-Qiou Hao, Jiou-Ping Wang, Rong-Xing Zhou, The Center of Diagnosis and Treatment of Infection Diseases of PLA, Affiliated Tangdu Hospital, Fourth Military Medical University, Xi'an 710038, Shaanxi Province, China
Supported by: the National Natural Science Foundation of China, No. 39800122.
Correspondence to: Zhi-Hua Feng, The Center of Diagnosis and Treatment of infection diseases of PLA, Tangdu Hospital, Fourth Military Medical University, Xi'an 710038, Shaanxi Province, China. email@example.com
Received: October 25, 2002 Revised: November 2, 2002 Accepted: November 19, 2002 Published online: February 15, 2003
AIM: To compare the biological characteristics of cultured mice bone marrow dendritic cell (BM-DC) in cultural media with peritoneum or with growth stimulating factor.
METHODS: DC progenitors of were isolated from bone marrow of Balb/C mice, and transformed into DC cell by culturing with secretary fluid of peritoneum, mGM-CSF, and mIL-4, respectively. The biological features and immunity of the DC cells were studied with mixed lymphocyte reaction, immunohistochemistry staining, light microscope and scanning electronic microscope.
RESULTS: After culturing for 6-8 days, the number of BM-DC (6x106) released from proliferating colony in media containing mGM-CSF and mIL-4 was higher than that in media containing peritoneum without significant difference. High level of expressed CD86 on BM-DC was demonstrated on day 7 in culture, MLR results indicated that the stimulating ability for proliferation of T cell was higher in media with mGM-CSF, mIL-4 than that with peritoneum, while the control media without stimulating factors showed no ability to stimulate the growth of T cells
CONCLUSION: These observations demonstrated that auto-peritoneum can substitute for cytokines in media and stimulate the growth of bone marrow-derived DCs, which possess the corresponding cellular biological features and cellular immunity.
Key Words: N/A
Citation: Wang QC, Feng ZH, Zhou YX, Nie QH, Hao CQ, Wang JP. Comparative research of dendritic cells cultured from mice bone marrow with different ways. Shijie Huaren Xiaohua Zazhi 2003; 11(2): 219-223
自1993年 Inaba et al用 GM-CSF 体外成功扩增 DC 以来, DC 的研究得到迅速发展. 骨髓中造血干细胞接种培养后, 在 GM-CSF、IL-4等 的作用下能向 DC 方向分化, 经历了一个由未成熟向成熟 DC 发育的过程. 早期未成熟 DC 的突起少, MHC 分子及共刺激分子 B7-1、B7-2的表达非常低, 无刺激 T 细胞增生的能力. 成熟后, 细胞突起增多, MHC 分子及共刺激分子的表达明显升高, 能强烈刺激 T 细胞的增生[29-32] . 这与我们观察到的实验结果完全符合, 即培养6-8 d 后 BM-DC 已基本成熟. 分离得到的 DC 前体细胞较少, DC 生长发育非常缓慢, 开始时仅少量的 DC 前体细胞在贴壁细胞的表面散在分布. 培养4 d, DC 前体细胞才开始出现小的集落. 至6-8 d, DC 前体细胞数量增多, 并出现一增生高峰, 有大量的半贴壁半悬浮集落生成; 少量悬浮细胞有小的突起. 此时DC 细胞表面分子 MHC class I, II和共刺激分子 B7-1, B7-2的表达都极低, 几乎没有刺激同种异体T细胞增生的能力. 这种 DC 的生物免疫特性与未成熟 DC 非常相似, 可能是诱导 T 细胞甚至凋亡的原因. 在培养中我们发现; 在7-8 d的增生高峰期, DC 有丰富的半贴壁集落生成; 并且从10 d起开始有部分细胞从集落中释放出来. 此时 DC 细胞表面分子表达有一定程度的升高, 以 MHC class II 和 B7-2为明显, 并且其刺激 T 细胞增生的能力也有增强.
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