中国组织工程研究

中国组织工程研究 ›› 2010, Vol. 14 ›› Issue (12): 2119-2122.doi: 10.3969/j.issn.1673-8225.2010.12.008

• 药物控释材料 drug delivery materials • 上一篇    下一篇

海藻糖深低温保存外周血造血干细胞的可行性

娄  琳，陆志刚   

  1. 南方医科大学珠江医院输血科，广东省广州市 510282
  • 出版日期:2010-03-19 发布日期:2010-03-19
  • 通讯作者: 陆志刚，硕士，教授，南方医科大学珠江医院输血科，广东省广州市 510282 gzluzhigang@126.com
  • 作者简介:娄 琳★，女，1983年生，山东省新泰市人，汉族，2010年南方医科大学毕业，硕士，医师，主要从事干细胞低温保存方面的研究。 llin2006@163.com

Feasibility of applying trehalose to cryopreserve peripheral blood stem cells

Lou Lin, Lu Zhi-gang   

  1. Department of Blood Bank, Zhujiang Hospital, Southern Medical University, Guangzhou  510282, Guangdong Province, China
  • Online:2010-03-19 Published:2010-03-19
  • Contact: Lu Zhi-gang, Master, Professor, Department of Blood Bank, Zhujiang Hospital, Guangzhou 510282, Guangdong Province, China gzluzhigang@126.com
  • About author:Lou Lin★, Master, Physician, Department of Blood Bank, Zhujiang Hospital, Southern Medical University, Guangzhou 510282, Guangdong Province, China llin2006@163.com

PDF

483

被引次数

7

摘要：

背景：二甲基亚砜是目前造血干细胞深低温保存的经典保护剂，但其对细胞和患者均有一定的毒副作用。海藻糖是一种稳定的无毒副作用的非还原性双糖，已被广泛应用于红细胞、血小板和胚胎等的冷冻保存中。
目的：探讨海藻糖作为低温保存造血干细胞保护剂的可行性。
方法：外周血造血干细胞经重组人集落刺激因子动员后，用血细胞分离机采集连续单个核细胞，分为0.5 mol/L海藻糖组、1.0 mol/L海藻糖组、对照组。采用程序降温法液氮保存，冻存7 d后取出，立即置于40 ℃水浴箱内复苏。锥虫蓝拒染法检测细胞存活率；采用甲基纤维素半固体培养体系进行集落培养，计数粒-巨噬细胞集落形成单位的回收率；采用CD34-PE/CD45-FITC双标法，流式细胞仪检测CD34+细胞回收率。
结果与结论：与对照组比较，0.5，1.0 mol/L海藻糖组细胞存活率、粒-巨噬细胞集落形成单位回收率、CD34+细胞回收率均明显升高(P < 0.001)，且0.5 mol/L海藻糖组升高幅度尤为显著(P < 0.001或P < 0.01)。证实海藻糖对于短期内低温冻存的外周血造血干细胞有一定保护作用，浓度为0.5 mol/L的海藻糖保护冻存的造血干细胞效果较佳。

关键词: 低温保存, 粒-巨噬细胞集落形成单位, 海藻糖, 浓度, 外周血造血干细胞, 生物材料

Abstract:

BACKGROUND: As the classical cryoprotectant for hematopoietic stem cells, dimethylsulfoxide has considerably toxicity for both the thawed cells and the patients. Trehalose is a stable and unharmful disaccharide, which has been widely used in the field of the cryopreseved red cells, platelet and organs.
OBJECTIVE: To explore the feasibility of trehalose used as cryoprotectant for hematopoietic stem cells.
METHODS: After recombinant colony stimulating factor mobilization, mononuclear cells were separated from hematopoietc stem cells and were divided into the control, 0.5 mol/L trehalose and 1.0 mol/L trehalose groups. The hematopoietc stem cells were auto-controlled programmed cryogenic system, then stored in liquid nitrogen for 1 week, followed by rapidly thawed in a 40 ℃ water bath. Cell survival rates were detected by trypan blue exclusion assay; the recovery rates of colony-forming unit granulocyte-macrophage and CD34+ cells were counted by CD34-PE/CD45-FITC double-staining methods and flow cytometry.
RESULTS AND CONCLUSION: Compared with the control group, cell survival, colony-forming unit granulocyte-macrophage and CD34+ cells recovery rate of the 0.5 and 1.0 mol/L trehalose groups were obviously increased, especially in the 0.5 mol/L trehalose group (P < 0.001 or P < 0.01). Trehalose has some cryoprotective potential for cryopreservation of hematopoietic stem cells, particularly, 0.5 mol/L exerts a superior cryoprotective potential. 

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