中国组织工程研究

中国组织工程研究 ›› 2019, Vol. 23 ›› Issue (29): 4681-4687.doi: 10.3969/j.issn.2095-4344.1814

• 脂肪干细胞 adipose-derived stem cells • 上一篇    下一篇

低氧条件下人脂肪间充质干细胞分泌多种细胞因子的水平

王  倩1，刘  羿2，张  雨3，杨印祥1，汪兆艳1，张乐平1，栾  佐1   

  1. 1中国人民解放军总医院第六医学中心儿科，北京市  100048；2中国人民解放军总医院第一医学中心输血科，北京市  100039；3首都医科大学附属北京妇产医院新生儿科，北京市  100026
  • 修回日期:2019-05-20 出版日期:2019-10-18 发布日期:2019-10-18
  • 通讯作者: 栾佐，硕士，主任医师，中国人民解放军总医院第六医学中心儿科，北京市 100048
  • 作者简介:王倩，女，1984年生，河北省安新县人，汉族，2009年河北北方学院毕业，主管技师，主要从事干细胞培养及相关课题研究。 并列第一作者：刘羿，1985年生，黑龙江省讷河市人，汉族，2009年河北北方学院毕业，主管技师，主要从事临床输血工作及相关课题研究。
  • 基金资助:

    全军后勤科研项目(CHJ14C022)，项目负责人：栾佐

The secretion levels of various cytokines in human adipose mesenchymal stem cells under hypoxia

Wang Qian1, Liu Yi2, Zhang Yu3, Yang Yinxiang1, Wang Zhaoyan1, Zhang Leping1, Luan Zuo1   

  1. 1Department of Pediatrics, The Sixth Medical Center of PLA General Hospital, Beijing 100048, China; 2Department of Pediatrics, The First Medical Center of PLA General Hospital, Beijing 100039, China; 3Department of Neonates, Beijing Obstetrics and Gynecology Hospital, Capital Medical University, Beijing 100026, China
  • Revised:2019-05-20 Online:2019-10-18 Published:2019-10-18
  • Contact: Luan Zuo, Master, Chief physician, Department of Pediatrics, The Sixth Medical Center of PLA General Hospital, Beijing 100048, China
  • About author:Wang Qian, Technologist-in- charge, Department of Pediatrics, The Sixth Medical Center of PLA General Hospital, Beijing 100048, China； Liu Yi, Technologist-in- charge, Department of Pediatrics, The First Medical Center of PLA General Hospital, Beijing 100039, China. Wang Qian and Liu Yi contributed equally to this work.
  • Supported by:

    the Military Logistics Research Project, No. CHJ14C022 (to LZ)

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摘要：

文章快速阅读：

文题释义：
脂肪干细胞：是从脂肪组织中分离得到的一种具有多向分化潜能的干细胞，它具有体内储存量大、获取容易、稳定增殖等特点。同时脂肪干细胞还具有强大的旁分泌作用，可分泌多种细胞因子，从而改善损伤或缺血性机体的生理和病理功能。
血管内皮生长因子：又称血管通透因子，具有促进血管通透性增加、细胞外基质变性、血管内皮细胞迁移、增殖和血管形成等作用。

 

摘要
背景：脂肪间充质干细胞来源于脂肪组织，可分泌多种细胞因子，成为治疗各类缺血性疾病的热点种子细胞之一。
目的：研究不同氧分压培养环境对人脂肪干细胞分泌细胞因子的影响，从而找出能使细胞因子高表达的最适氧分压。
方法：利用酶消化法从人脂肪组织中分离脂肪干细胞，通过细胞形态学特征、体外分化功能及免疫表型对分离培养的细胞进行鉴定。人脂肪干细胞分别于3%，5%，21%的氧分压环境中培养48 h，采用酶联免疫吸附实验检测血管内皮生长因子、脑源性神经营养因子、类胰岛素一号增长因子的分泌水平。
结果与结论：①人脂肪干细胞呈长梭形或多角形，胞体丰满，胞浆均匀，呈漩涡状生长，体外诱导可进行成脂、成骨和成软骨分化；②流式细胞仪检测结果显示细胞高表达间充质表面标志物CD29、CD73、CD90和CD105，而几乎不表达造血干细胞表面标志物CD34和CD45，阴性表达HLA-DR；③人脂肪干细胞在低氧条件下(3%，5%氧分压)血管内皮生长因子的分泌量为常氧条件下的2.3倍(P < 0.05)和1.5倍(P < 0.05)；低氧条件下(3%，5%氧分压)类胰岛素一号增长因子的分泌量分别为常氧条件下的2.4倍(P < 0.05)和1.5倍(P < 0.05)；低氧条件下(3%，5%氧分压)脑源性神经营养因子的分泌量与常氧条件下差异无显著性意义(P > 0.05)；④结果表明，人脂肪干细胞在低氧环境中可显著提高血管内皮生长因子和类胰岛素一号增长因子的分泌量，而脑源性神经营养因子的分泌量并未受到影响。


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
ORCID: 0000-0002-4638-4367(王倩)

关键词: 低氧, 人脂肪干细胞, 血管内皮生长因子, 类胰岛素一号增长因子, 脑源性神经营养因子, 氧分压

Abstract:

BACKGROUND: Mesenchymal stem cells derived from adipose tissue can secrete a variety of cytokines. Therefore, these cells have become hot seed cells for treating various ischemic diseases.
OBJECTIVE: To explore the effects of different oxygen partial pressures on the secretion of cytokines from human adipose mesenchymal stem cells, and to select the optimal oxygen partial pressures under which cytokines can highly express.
METHODS: Adipose mesenchymal stem cells were isolated from human adipose tissue by enzymatic digestion, and were identified by morphological characteristics, differentiation function and immunophenotype. Human adipose mesenchymal stem cells were cultured at 3%, 5%, and 21% partial oxygen pressures for 48 hours, respectively. The secretion levels of vascular endothelial growth factor, brain-derived neurotrophic factor, and insulin-like growth factor-1 in the cells were detected by enzyme-linked immunosorbent assay.
RESULTS AND CONCLUSION: Human adipose mesenchymal stem cells grew spirally with a long fusiform or polygonal shape, plump cell body, and uniform cytoplasm. They could be induced for adipogenic, osteogenic and chondrogenic differentiation in vitro. The cultured cells highly expressed mesenchymal surface markers CD29, CD73, CD90, and CD105, scarcely expressed hematopoietic stem cell surface markers CD34 and CD45, and negatively expressed HLA-DR. The level of vascular endothelial growth factors secreted by human adipose mesenchymal stem cells under 3% or 5% partial pressure of oxygen (hypoxia) was 2.3 times (P < 0.05) or 1.5 times (P < 0.05) as high as that under normoxia, respectively. The secretion level of insulin-like growth factor-1 under 3% or 5% partial pressure of oxygen (hypoxia) was 2.4 times (P < 0.05) or 1.5 times (P < 0.05)higher than that under normoxia, respectively. However, there was no significant difference in the secretion levels of brain-derived neurotrophic factor between hypoxic and normoxic conditions (P > 0.05). Therefore, human adipose mesenchymal stem cells significantly increase the secretion of vascular endothelial growth factor and insulin-like growth factor-1 under hypoxic conditions, while the secretion of brain-derived neurotrophic factor dose not change under hypoxic conditions.

Key words: hypoxia, human adipose stem cells, vascular endothelial growth factor, insulin-like growth factor-1, brain-derived neurotrophic factor, oxygen partial pressure

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