Freeze-thawing procedures have no influence on the phenotypic and functional development of dendritic cells generated from peripheral blood CD14+ monocytes
Freeze-thawing procedures have no influence on the phenotypic and functional development of dendritic cells generated from peripheral blood CD14+ monocytes
复制标题
DOI：
10.1097/00002371-200401000-00003
复制
发表时间：
2004-01-01
影响因子：
3.9
通讯作者：
Kakizoe, T
中科院分区：
文献类型：
--
作者：
Hori, S;Heike, Y;Kakizoe, T
冻存对树突状细胞（DC）生物学活性的潜在影响知之甚少。在这项研究中，我们研究了冻融对从粒细胞集落刺激因子（G-CSF）动员的外周血CD 14（+）细胞获得的人DC的表型和功能发育的影响。CD 14（+）细胞在冻融后立即用粒细胞-巨噬细胞CSF和白细胞介素-4培养9天，然后加入肿瘤坏死因子-α再培养3天。对于新鲜和冻融的单核细胞，在相同的条件下检查培养第6天收获的未成熟DC和培养第9天收获的成熟DC。比较细胞的1）抗原内吞和趋化性迁移能力（未成熟DC），和2）同种异体混合淋巴细胞反应和抗原特异性细胞毒性T淋巴细胞应答（成熟DC）。冻融不影响任何发育阶段的DC的活力或随后的成熟。此外，从新鲜或冻存/解冻细胞产生的细胞之间在表型或功能上基本上没有观察到差异。尽管该研究设计仅限于使用胎牛血清，但观察结果仍然表明冻融不影响任何成熟阶段的DC的活力、表型、随后的成熟或功能。
Little is known about the potential influence of cryopreservation on the biologic activities of dendritic cells (DCs). In this study, we examined the effects of freeze-thawing on the phenotypic and functional development of human DCs obtained from granulocyte colony-stimulating factor (G-CSF)-mobilized peripheral blood CD14(+) cells. CD14(+) cells were cultured, immediately or after freeze-thawing, with granulocyte-macrophage CSF and interleukin-4 for 9 days, and then with added tumor necrosis factor-alpha for another 3 days. For both fresh and freeze-thawed monocytes, immature DCs harvested on day 6 and mature DCs harvested on day 9 of culture were examined under the same conditions. Cells were compared with regard to their 1) capacities for antigen endocytosis and chemotactic migration (immature DCs), and 2) allogeneic mixed lymphocyte reaction and antigen-specific cytotoxic T lymphocyte responses (mature DCs). Freeze-thawing did not affect the viability or subsequent maturation of DCs at any stage of development. Furthermore, essentially no difference was observed in phenotype or function between cells generated from fresh or cryopreserved/thawed cells. Although this study design was limited with the use of fetal bovine serum, the observation still suggests that freeze-thawing does not affect viability, phenotype, subsequent maturation, or functions of DCs at any stage of maturation.