中国组织工程研究

中国组织工程研究 ›› 2010, Vol. 14 ›› Issue (45): 8399-8402.doi: 10.3969/j.issn.1673-8225.2010.45.010

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

锌对人脐带血间充质干细胞DNA和蛋白质含量及增殖周期的影响

姚素艳1，杨依勇2，秦书俭2，郑德宇2   

  1. 辽宁医学院，1病理生理学教研室，2解剖学教研室，辽宁省锦州市  121001
  • 出版日期:2010-11-05 发布日期:2010-11-05
  • 通讯作者: 郑德宇，副教授，辽宁医学院解剖学教研室，辽宁省锦州市 121001 zheng_deyu2000@yahoo.com.cn
  • 作者简介:姚素艳★，女，1971年生，辽宁省兴城市人，满族，2005年辽宁医学院毕业，硕士，副教授，主要从事神经退行性变和骨组织工程方面的研究。 ysyzdy@yahoo.com.cn
  • 基金资助:

    课题受到辽宁省教育厅创新团队(2006T06110)的支持，课题名“同种异体种子细胞的抗原性对大鼠桡骨缺损修复的影响”。

Effects of zinc on DNA and protein contents as well as reproductive cycle of human umbilical cord blood-derived mesenchymal stem cells

Yao Su-yan1, Yang Yi-yong2, Qin Shu-jian2, Zheng De-yu2   

  1. 1 Department of Pathophysiology, 2 Department of Anatomy, Liaoning Medical University, Jinzhou  121001, Liaoning Province, China
  • Online:2010-11-05 Published:2010-11-05
  • Contact: Zheng De-yu, Associate professor, Department of Anatomy, Liaoning Medical University, Jinzhou 121001, Liaoning Province, China zheng_deyu2000@yahoo.com.cn
  • About author:Yao Su-yan★, Master, Associate professor, Department of Pathophysiology, Liaoning Medical University, Jinzhou 121001, Liaoning Province, China ysyzdy@yahoo.com.cn
  • Supported by:

    the Innovation Team of Education Department of Liaoning Province, No. 2006T06110*

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摘要：

背景：锌通过直接和间接作用对DNA聚合酶和RNA聚合酶产生影响，在细胞的DNA复制、RNA转录、增殖、分化等活动中起重要作用。有研究表明适宜浓度的锌具有促进神经元和成骨细胞增殖的作用。
目的：拟通过流式细胞技术观察锌对脐血间充质干细胞细胞周期、DNA和蛋白质含量的影响，旨在探讨适宜浓度锌对脐血间充质干细胞的干预作用。
方法：密度梯度离心法分离培养人脐带血间充质干细胞，取传3代细胞，对照组向细胞中加入含胎牛血清的DMEM/F12培养液，锌处理组向上述培养液中加入硫酸锌，使锌终浓度分别达到0.5，1.5，2.5，3.5，4.5，5.5 mg/L，培养21 d。观察细胞表面抗原的表达，MTT法检测细胞活性，流式细胞仪测定细胞DNA含量、蛋白质含量和增殖周期。
结果与结论：脐带血间充质干细胞表面抗原CD29和CD44呈阳性表达。锌对脐带血间充质干细胞活性的促进作用呈量效关系，0.5~4.5 mg/L锌处理组细胞活性显著高于对照组(P < 0.01)，尤以质量浓度为2.5 mg/L时最为明显。培养7，14，21 d与对照组相比，2.5 mg/L锌处理组脐带血间充质干细胞DNA含量、蛋白质含量均显著升高(P < 0.01)；细胞增殖指数、S期和G2+M期细胞百分率均明显升高，G0/G1期细胞百分率则显著下降(P < 0.01)。提示0.5~4.5 mg/L锌能促进脐带血间充质干细胞的增殖、DNA复制和蛋白质合成，且2.5 mg/L为最佳质量浓度。

关键词: 锌, 脐血间充质干细胞, 增殖, DNA, 蛋白质

Abstract:

BACKGROUND: Zinc has effects on DNA polymerase and RNA polymerase by direct and indirect action, and plays important roles in DNA duplication, RNA transcription, proliferation and differentiation of cells. Some studies have verified that zinc at a suitable concentration has effects on promoting the proliferation of neurons and osteoblasts.
OBJECTIVE: To explore effects of zinc on DNA and protein content as well as reproductive cycle of human umbilical cord blood-derived mesenchymal stem cells (UCB-MSCs) using flow cytometry, and to investigate zinc at a suitable concentration effects on UCB-MSCs.
METHODS: UCB-MSCs were isolated and cultured in vitro by the density gradient centrifugation. At the third passage, BMSCs in the control group were incubated with DMEM/F12 containing fetal bovine serum. BMSCs in the zinc group were incubated in above-mentioned DMEM, supplemented with zinc sulfate. The final concentration of zinc was 0.5, 1.5, 2.5, 3.5, 4.5 and 5.5 mg/L. BMSCs were incubated for 21 days. Cell surface antigen expression was observed. MTT assay was used to detect cell activity. Flow cytometry was utilized to measure DNA content, protein content and reproductive cycle.
RESULTS AND CONCLUSION: Cells were positive for CD29 and CD44. Zinc had dose-effect relationship on promotion of BMSC activity. Cell activity was significantly higher in the 0.5-4.5 mg/L zinc group compared with the control group (P < 0.01), especially at 2.5 mg/L. At 7, 14 and 21 days, BMSC DNA content and protein content significantly increased in the 2.5 mg/L zinc group compared with the control group (P < 0.01). Cell percentage in the S and G2+M phases and cell proliferation index significantly increased, but cell percentage in the G0/G1 phases decreased significantly (P < 0.01). These indicate that zinc (0.5-4.5 mg/L) has the effect of promoting the rat BMSC proliferation, DNA reproduction and protein synthesis, and the 2.5 mg/L is the optimal concentration.

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