当归多糖对脐血造血细胞冷冻损伤的可恢复性研究

Effect of APS on recoverable ability from cryopreservation damage of UCB hematopoietic cells

目的探讨当归多糖(APS)是否具有恢复脐血造血细胞冷冻损伤的作用及与造血生长因子(HGFs)联合促进冷冻复苏后脐血单个核细胞(MNC)体外扩增的效应。方法将冷冻30d的脐血MNC复苏后,立即在常规培养体系中分别加入APS0、50、100、200、400μg/mL,或同时加入HGFs组合,培养24h或14d,采用MNC计数,台盼蓝拒染实验、MTT法、CFU-Mix集落形成实验以及流式细胞术计数CD34+细胞等方法分别观察APS促进冷冻损伤的脐血造血细胞恢复的能力以及APS联合HGFs对脐血造血细胞的扩增能力。结果冷冻复苏后加入一定质量浓度的APS可使脐血MNC数量与台盼蓝拒染率明显增加,造血细胞的增殖能力显著提高,CFU-Mix集落产率明显提高(P<0.05),且能明显提高冻存脐血MNC中CD34+细胞率(P<0.05);一定质量浓度的APS联合HGFs可显著提高冷冻复苏后的脐血MNC扩增倍数及CFU-Mix集落产率(P<0.05);其作用无明显量效关系。结论APS能提高冷冻复苏后脐血造血细胞的活力、增殖能力以及CD34+细胞率,可促进脐血造血细胞冷冻损伤的恢复;与HGFs联合可促进冷冻复苏后的脐血MNC体外扩增。

Objective To study the effect of angelica polysaecharides （APS） on recoverable ability from cryopreservation damage of umbilical cord blood （UCB） hematopoietic cells and APS with hematopoietic growth factors （HGFs） on in vitro expansion of UCB mononuclear cells （MNC） after thawing. Methods Thawed UCB MNC were cultured 24 h or 14 d with various concentration of APS （0, 50, 100, 200, and 400 ~g/mL） or with APS and HGFs. The MNC counting assay, typan blue exclusion assay, colorimetric MTT assay for cell proliferation, CFU-Mix colong-forming assay, and flow cytometry for CD34＋ cell rate were detected. Results After adding certain concentration APS in thawed UCB cells, the number of MNC, the rate of trypan blue exclusion, the proliferation of MNC, and the colony production of CFU-Mix were significantly enhanced （P〈0.05）. The percentage of CD34＋ cells in UCB was markedly higher than that of control group （P〈0. 05）. The certain concentration APS combined with HGFs could significantly raise the expansion fold and the colony production of CFU-Mix of thawed UCB hematopoietic cells （P〈0. 05）. There is no dose-response relation. Conclusion APS could raise the postfreezing recoverable ability of UCB hematopoietic cells. And APS combined with HGFs could promote the in vitro expansion of hematopoietic cells after thawed.