中国组织工程研究 ›› 2014, Vol. 18 ›› Issue (6): 962-967.doi: 10.3969/j.issn.2095-4344.2014.06.023

• 干细胞培养与分化 stem cell culture and differentiation • 上一篇    下一篇

围术期心血管危险因素对骨髓祖细胞的影响

张 林,高长青,王 嵘,李利兵,成 楠,姚名辉   

  1. 解放军心脏外科研究所,解放军总医院心血管外科,北京市 100853
  • 修回日期:2013-12-03 出版日期:2014-02-05 发布日期:2014-02-05
  • 通讯作者: 高长青,博士,主任医师,解放军心脏外科研究所,解放军总医院心血管外科,北京市 100853
  • 作者简介:张林,男,1978年生,宁夏回族自治区灵武市人,汉族,2011年解放军医学院毕业,博士,主治医师,主要从事成人缺血性心脏病的研究。
  • 基金资助:

     the National High-Technology Research and Development Plan of China (863 Program), No. 2006AA02A104

Impact of perioperative cardiovascular risk factors on bone marrow progenitor cells

Zhang Lin, Gao Chang-qing, Wang Rong, Li Li-bing, Cheng Nan, Yao Ming-hui   

  1. Department of Cardiovascular Surgery, PLA General Hospital, PLA Institute of Cardiac Surgery, Beijing 100853, China
  • Revised:2013-12-03 Online:2014-02-05 Published:2014-02-05
  • Contact: Gao Chang-qing, M.D., Chief physician, Department of Cardiovascular Surgery, PLA General Hospital, PLA Institute of Cardiac Surgery, Beijing 100853, China
  • About author:Zhang Lin, M.D., Attending physician, Department of Cardiovascular Surgery, PLA General Hospital, PLA Institute of Cardiac Surgery, Beijing 100853, China
  • Supported by:

    “十一五”国家高技术研究发展计划(863计划)(2006AA02A104),课题名称:冠状动脉旁路移植术联合骨髓单个核细胞移植治疗重症冠心病心肌梗死的临床研究

摘要:

背景:移植自体骨髓干细胞治疗缺血性心脏病已进行了10余年的临床试验,但试验结果在不同的患者中存在差异。因此,有必要鉴定哪些心血管病患者的危险因素影响骨髓干细胞的水平和功能。

目的:观察冠心病患者围术期危险因素对骨髓祖细胞数量及功能的影响。
方法:选择44例拟行冠状动脉旁路移植的冠心病患者,采集实验室和临床资料;术中经胸骨穿刺采集骨髓,应用Ficoll淋巴细胞分离液密度梯度离心法分离骨髓单个核细胞,计数并应用锥虫蓝拒染法检测其活性;应用流式细胞仪分析检测CD34+、CD133+和CD34+CD133+细胞的水平;应用集落形成试验和细胞迁移试验评价骨髓祖细胞功能。
结果与结论:术中经胸骨抽取20 mL骨髓可获得(10-89)×106个骨髓单个核细胞,活性在95%以上,等量的骨髓血获得的骨髓单个核细胞的量与患者年龄之间存在明显负相关关系(n=44,r=-0.788,P=0.001);流式细胞仪检测CD34+细胞占(0.94±0.39)%,CD133+细胞占(0.46±0.28)%,CD34+CD133+细胞占(0.53±0.26)%;糖尿病患者骨髓CD34+和CD133+细胞水平明显低于非糖尿病患者;高龄、女性和心功能较差与骨髓祖细胞集落形成能力降低有关;CD34+细胞水平与骨髓单个核细胞的迁移能力存在明显的正相关。结果表明经胸骨应用密度梯度离心法可获得足够数量的骨髓单个核细胞作为缺血性心脏病治疗的供体细胞,年龄、性别、糖尿病、心功能与骨髓单个核细胞数量和功能有关。

中国组织工程研究杂志出版内容重点:干细胞;骨髓干细胞;造血干细胞;脂肪干细胞;肿瘤干细胞;胚胎干细胞;脐带脐血干细胞;干细胞诱导;干细胞分化;组织工程


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关键词: 干细胞, 骨髓干细胞, 骨髓祖细胞, 冠心病, 冠状动脉旁路移植术, 细胞治疗, 863项目

Abstract:

BACKGROUND: Cell therapy by the implantation of autologous bone marrow cells has been used for the treatment of ischemic heart diseases in clinical trials for decade. However, as the outcomes of cell transplantation obviously vary among patients, it is essential to identify the risk factors that may influence the level and function of progenitor cells in bone marrow, in order to identify the patients who would benefit the most from this treatment.

OBJECTIVE: To observe the impact of perioperative cardiovascular risk factors on number and function of bone marrow progenitor cells from patients undergoing coronary artery bypass grafting surgery.
METHODS: We collected clinical and laboratory data from 44 patients scheduled to undergo sternotomy for coronary artery bypass grafting procedures. Bone marrow was aspirated from the sternum during the operation and bone marrow mononuclear cells were isolated by density centrifugation with Ficoll lymphoprep and then detected using trypan blue exclusion method. Levels of progenitor cells in bone marrow were evaluated using flow cytometry. Function of bone marrow progenitor cells were assessed by clonogenic and migration assays.
RESULTS AND CONCLUSION: We assessed the number of bone marrow mononuclear cells out of 20 mL bone marrow in duplicate samples from patients with coronary heart disease scheduled for coronary artery bypass grafting that was (10-89)×106 cells with over 95% activity. A negative correlation was observed between the number of bone marrow mononuclear cells and the age (n=44, r=-0.788, P=0.001). Levels of CD34+, CD133+, and CD34+CD133+ cells in bone marrow mononuclear cells was (0.94±0.39)%, (0.46±0.28)%, and (0.53±0.26)%. Levels of CD34+ cells and CD133+ cells in patients with diabetes were significantly lower than those in patients without diabetes. Female, advanced age and poor heart function were related with reduced colony-forming ability of progenitor cells. A positive correlation was observed between level of CD34+ cells and migration ability of bone marrow mononuclear cells. The results show that by density gradient centrifugation, we can harvest a sufficient number of bone marrow mononuclear cells in the treatment for ischemic heart disease. Age, gender, diabetes, heart function are correlated with bone marrow mononuclear cell number and functions.

中国组织工程研究杂志出版内容重点:干细胞;骨髓干细胞;造血干细胞;脂肪干细胞;肿瘤干细胞;胚胎干细胞;脐带脐血干细胞;干细胞诱导;干细胞分化;组织工程


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Key words: coronary disease, heart diseases, myeloid progenitor cells, coronary artery bypass, off-pump

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