基础医学与临床 ›› 2017, Vol. 37 ›› Issue (1): 13-18.

• 研究论文 • 上一篇    下一篇

持续低氧预处理促进兔尿源性干细胞增殖和减少凋亡

马文军1,储加强2,陶立1,龚梦嘉3,毕杨3,魏光辉1,张元原1   

  1. 1. 重庆医科大学附属儿童医院
    2. 重庆医科大学
    3. 重庆医科大学附属儿童医院儿研所干细胞实验室//儿童发育疾病研究教育部重点实验室//重庆市干细胞治疗工程技术研究中心
  • 收稿日期:2016-03-30 修回日期:2016-05-28 出版日期:2017-01-05 发布日期:2016-12-30
  • 通讯作者: 张元原 E-mail:yyzhang2005@gmail.com
  • 基金资助:
    国家自然科学基金资助项目

Sustained hypoxia promotes the proliferation and reduces apoptosis of rabbit urine derived stem cells

  • Received:2016-03-30 Revised:2016-05-28 Online:2017-01-05 Published:2016-12-30

摘要: 目的:探讨持续低氧(3%O2)环境对兔尿源性干细胞(rabbit urine-derived stem cells,rUSCs)增殖、凋亡及分泌功能的影响;方法: rUSCs分别置于3% O2和20% O2条件下培养,连续观察其形态学变化,绘制细胞增殖曲线;检测其分泌、周期及凋亡变化;结果:rUSCs原代培养约第6d便可见单个“米粒样”细胞,不同氧浓度培养后发现常氧组细胞为“长梭形”,低氧组向“鹅卵石”样改变;两组细胞增殖曲线均呈“S”形,但低氧组增殖速度较快(P<0.01);低氧组较常氧组相比,血管营养因子表达量均有上调且表达因子种类增加;常氧组G0/G1期细胞数为(91.73%±1.35%),明显高于低氧组的(71.82%±8.86%)(P<0.05);常氧组S期细胞数为(3.48%±0.73%),显著低于低氧组的(18.77%±3.87%)(P<0.01);流式常氧组凋亡率为(3.24%±0.38%),明显高于低氧组的(0.43%±0.05%)(P<0.001);Hoechst结果常氧组凋亡率为(4.27%± 0.48%),明显高于低氧组的(1.37%± 0.33%)(P<0.01)。结论:持续性低氧(3% O2)环境可促进rUSCs细胞的增殖,减少细胞凋亡,增加其分泌功能。

关键词: 兔尿源性干细胞, 低氧, 增殖, 凋亡

Abstract: Objective To investigate the effects of hypoxia on the cell proliferation, apoptosis and secretion of rabbit urine derived stem cells (rUSCs);Methods rUSCs were isolated and cultured with oxygen concentration at 20 % and 3 %, respectively. To observe the morphological changes of rUSCs in different culture conditions continuously; The growth curves under different culture conditions were drawn respectively;Antibody detection of factor secretion was performed;The cell cycle and the apoptosis were analyzed by flow cytometry. Results Single, small, compact “rice-grain” like cells were observed on day 6, after cultured in different oxygen concentration,the cells in normoxia group appeared like "spindle", the cells in hypoxia group changed into " pebble" like; The growth curves of two groups showed a "S" shape, but the cells in hypoxia group had a faster growth speed compared with normoxia group (P <0.01 resp.); Compared with normoxia group, the paracrine effect in hypoxia group had an increased expression level , and an increased variety ; Less cells were arrested at G0/G1 phase under hypoxia as compared with that in normoxia group(71.82%±8.86% and 91.73%±1.35%, resp.) (P<0.05), while the cell numbers at S phase increased in hypoxia group are comparied with that of normoxia group(18.77%±3.87% and 3.48%±0.73%, resp.) (P<0.01); The apoptosis rate of normoxia group(3.24%±0.38% ) was significantly higher than that of hypoxia group by flow cytometry (0.43%±0.05%) (P<0.001); and the apoptosis rate of normoxia group(4.27%± 0.48% ) was significantly higher than that of hypoxia group by Hoechst(1.37%± 0.33%)(P<0.01). Conclusion It is demonstrated that culture at 3% O2 could enhance cell proliferation and reduce apoptosis of rUSCs.

Key words: rabbit urine derived stem cells, hypoxia, cell proliferation, cell apoptosis

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