G-CSF 动员的外周血干细胞悬液冷冻保存后诱导培养树突状细胞的研究

目的：研究粒细胞集落刺激因子（G-CSF） 动员的外周血干细胞（PBSC）悬液中单个核细胞(MNC)冷冻保存后经不同诱导途径培养的树突状细胞(DC)的特性，探讨PBSC悬液冷冻保存后诱导培养DC的方法。 方法： 用两种保护液-80 ℃冷冻保存PBSC悬液，分析复苏MNC中CD34+细胞、CD14+细胞、CD14+PI+细胞，并分两种方法诱导培养复苏MNC: 贴壁细胞经粒细胞单核细胞集落刺激因子（GM-CSF）、白细胞介素4（IL-4）培养2周，培养结束前加入肿瘤坏死因子α（TNF-α）、布雷菲德菌素（BFA）；非贴壁细胞经酪氨酸激酶3配体（FL）、干细胞因子（SCF）、GM-CSF、IL-4培养1周, 再按前1种方法继续培养2周。培养结束后, 流式细胞仪分析DC特性。 结果： 两组复苏MNC与新鲜MNC比较：CD34+细胞含量均无明显差异(P>0.05)；而CD14+细胞少，CD14+PI+细胞百分率高（P<0.01）；贴壁细胞少，贴壁细胞诱导培养DC效果不佳；非贴壁细胞扩增诱导均能获得大量成熟的激活的能够分泌IL-12(p40)的髓系DC。 结论： PBSC悬液冷冻保存后能用于培养髓系DC，但培养方法应以扩增诱导为主。

Dendritic cells induced from culture of cryopreservated G-CSF-mobilized peripheral blood stem cell harvests

AIM: To find the way of culturing the dendritic cells (DC) from the cryopreservated G - CSF - mobilized peripheral blood stem cell(PBSC) harvests. METHODS: The non - adherent or adherent cells from the fresh or refreshed PBSC harvests were induced to DCs separately. The amount of CD34 cells, CD14 PI cells in the PBSC harvests and the characters of DCs were analyzed by flow cytometry. RESULTS: The DCs cultured from the non-adherent cells were similar to those from the fresh groups in the number and the characters. The amount of CD34 cells was also similar between them, but CD14 PI cells were higher in the refreshed groups than that in the fresh groups. The adherent cells from the refreshed group were too few to be induced to DCs. CONCLUSION: The refreshed PBSC harvests could be induced to the matured activated myloid DCs cultured by the routine of CD34 cells rather than by the way of monocytes.