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人脐带间充质干细胞中神经营养因子的表达
  • ISSN号:1671-8925
  • 期刊名称:中华神经医学杂志
  • 时间:2012
  • 页码:438-442
  • 分类:R329.2[医药卫生—人体解剖和组织胚胎学;医药卫生—基础医学]
  • 作者机构:[1]北京海淀医院神经外科,北京100080, [2]解放军总医院骨科研究所,北京100853
  • 相关基金:国家自然科学基金(31100696);国家科技支撑计划(2009BAl87802)
  • 相关项目:神经组织来源的细胞外基质支架复合人脐带Wharton胶间充质干细胞构建组织工程神经
中文摘要:

目的通过检测人脐带间充质干细胞(hUE.MSCs)的神经营养因子谱,以进一步认识hUC—MSCs通过分泌神经营养因子治疗神经损伤的作用。方法组织块培养法分离培养hUC-MSCs.流式细胞仪鉴定干细胞表面标记物,Raybio人细胞因子抗体芯片检测hUC-MSCs神经营养因子表达情况,RT—PCR检测hUC.MSCs神经营养因子的基因表达,ELISA法定量检测hUC.MSCs上清液中分泌的脑源性神经营养因子(BDNF)、胶质细胞源性神经营养因子(GDNF)、肝细胞生长因子(HGF)、神经营养因子3(NT.3)含量。结果流式细胞分析结果显示hUC—MSCs表面标记物CD29、CD44、CD90阳性(94.05%、90.75%、98.12%),CD34、CD45阴性(3.09%、0.80%)。Raybio人细胞因子抗体芯片结果显示,与神经营养相关的因子BDNF、表皮生长因子(EGF)、GDNF、HGF、NT-3、血管内皮细胞生长因子(VEGF)、胰岛素样生长因子.I(IGF一1)、神经营养因子4(NT.4)在hUC.MSCs中均有表达。RT.PCR检验也证实,GnJ、VEGF、神经生长因子(NGF)、HGF、GDNF、NT-4、BDⅣF和Ⅳ卜3基因在hUC—MSCs表达。ELISA检测细胞上清液中BDNF、GDNF、HGF、NT.3的含量分别为(475.20±32.22)pg/mL、(82.33±3.39)pg/mL、(704.50±12.86)pg/mL、(230.41±16.66)pg/mL,分别与空白对照组相比,差异均有统计学意义(P〈0.05)。结论hUC.MSCs富含神经营养因子,是一种理想的用于神经损伤治疗的种子细胞。

英文摘要:

Objective To observe the profile of neurotrophic factors in human umbilical cord-derived mesenchymal stromal cells (hUC-MSCs) with Raybio antibody chip system, and provide a new cell seed source for spine cord injury or peripheral nerve regeneration. Methods The hUC-MSCs were cultured by tissue culture. Characteristics of cell phenotype were confirmed by flow cytometry. Raybio human cytokine antibody array and RT-PCR were employed to detect the neurotropic factors in hUC-MSCs. ELISA was used to determine the contents of growth factors (brain-derived neurotrophic factor [BDNF], glial cell line-derived neurotrophic factor [GDNF], hepatocyte growth factor [HGF], neurotrophin-3 [NT-3]) secreted by hUC-MSCs. Results The hUC-MSCs showed positive results for CD29 (94.05%), CD44 (90.75%) and CD90 (98.12%), and negative results for CD34 (3.09%) and CD45 (0.80%). Array results showed that hUC-MSCs expressed BDNF, epidermal growth factor (EGF), GDNF, HGF and NT-3, vascular endothelial growth factor (VEGF), insulin-like growth factor 1 (IGF-1) and NT-4. The gene expressions of IGF-1, VEGF, nerve growth factor (NGF), HGF, GDNF, NT-4, BDNF and NT-3 were further verified by RT-PCR. ELISA results showed that the contents of BDNF, GDNF, HGF and NT-3 in the conditioned medium were (475.20±32.22) pg/mL, (82.33±3.39) pg/mL, (704.50±12.86) pg/mL, and (230.41 ±16.66) pg/mL, respectively. These were obviously higher than that in the controlgroup. Conclusion The hUC-MSCs, containing plenty of growth factors, especially neurotrophic factors, may be promising seed cells for spine cord injury and nerve regeneration.

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期刊信息
  • 《中华神经医学杂志》
  • 北大核心期刊(2011版)
  • 主管单位:中华人民共和国卫生部
  • 主办单位:中华医学会
  • 主编:
  • 地址:广东省广州市工业大道中253号
  • 邮编:510282
  • 邮箱:Journal@126.com
  • 电话:020-61643273
  • 国际标准刊号:ISSN:1671-8925
  • 国内统一刊号:ISSN:11-5354/R
  • 邮发代号:46-251
  • 获奖情况:
  • 国内外数据库收录:
  • 美国化学文摘(网络版),中国中国科技核心期刊,中国北大核心期刊(2008版),中国北大核心期刊(2011版),中国北大核心期刊(2014版)
  • 被引量:16608