中国组织工程研究

中国组织工程研究 ›› 2010, Vol. 14 ›› Issue (32): 6001-6005.doi: 10.3969/j.issn.1673-8225.2010.32.024

• 干细胞基础实验 basic experiments of stem cells • 上一篇    下一篇

血管内皮细胞和脂肪干细胞体外联合培养细胞的增殖

王福科1，刘 流1，何晓光1，赵德萍2，袁瑞红1，代晓明1，李逸松1   

  1. 昆明医学院第一附属医院，1头颈外科，2实验中心，云南省昆明市 650032
  • 出版日期:2010-08-06 发布日期:2010-08-06
  • 通讯作者: 刘流，博士，教授，博士生导师，昆明医学院第一附属医院头颈外科，云南省昆明市 650032 liuliu3939@126.com
  • 作者简介:王福科☆，男，1975年生，河南省焦作市人，汉族，2009年昆明医学院毕业，博士，讲师，主要从事骨组织工程方面的研究。 wfk.04@126.com
  • 基金资助:

    国家自然科学基金(30960388)，课题名称“hVECs联合培养体系中Bmi-1,Runx2基因变化与hBMSCs的增殖和成骨分化研究”；云南科技厅联合专项基金资助项目(2007C0007R)，课题名称“脐血内皮细胞与骨髓间充质干细胞联合培养移植修复颌骨缺损”；昆明医学院博士创新基金项目(KM2007D03)，课题名称“异源间充质干细胞对新型仿生骨衍生材料移植的免疫调节作用研究”。

Proliferation of vascular endothelial cells and adipose tissue-derived stem cells under in vitro coculture

Wang Fu-ke1, Liu Liu1, He Xiao-guang1, Zhao De-ping2, Yuan Rui-hong1, Dai Xiao-ming1, Li Yi-song1   

  1. 1 Department of Head and Neck Surgery, 2 Experimental Center, First Affiliated Hospital of Kunming Medical College, Kunming  650032, Yunnan Province, China
  • Online:2010-08-06 Published:2010-08-06
  • Contact: Liu Liu, Doctor, Professor, Doctoral supervisor, Department of Head and Neck Surgery, First Affiliated Hospital of Kunming Medical College, Kunming 650032, Yunnan Province, China liuliu3939@126.com
  • About author:Wang Fu-ke☆, Doctor, Lecturer, Department of Head and Neck Surgery, First Affiliated Hospital of Kunming Medical College, Kunming 650032, Yunnan Province, China wfk.04@126.com
  • Supported by:

    the National Natural Science Foundation of China, No. 30960388*; the Associated Specific Foundation Program of Department of Science and Technology of Yunnan Province, No. 2007C0007R*; the Doctor Innovation Foundation Program of Kunming Medical College, No. KM2007D03*

PDF

291

被引次数

4

摘要：

背景：血管内皮细胞可有力支持脂肪干细胞向成骨方向转变，还可以加快血管的发生，为干细胞成骨提供营养支持。
目的：探讨血管内皮细胞和脂肪干细胞体外联合培养对混合细胞增殖的影响。
方法：设立单纯血管内皮细胞培养组、脂肪干细胞与血管内皮细胞按3∶1，1∶3，1∶1混合培养组、单纯脂肪干细胞培养组，加入含胎牛血清的L-DMEM，置于37 ℃、体积分数为5%的CO2饱和湿度培养箱中。倒置显微镜下观察细胞形态变化，改良MTT法绘制细胞生长曲线。
结果与结论：脂肪干细胞与血管内皮细胞按3∶1，1∶1混合培养14 d，细胞之间出现许多突触连接，部分细胞融合成团块状，其余培养方式均未见细胞团出现。随培养时间的延长，各培养方式下细胞吸光度值均逐渐升高，单纯脂肪干细胞培养、脂肪干细胞与血管内皮细胞按3∶1，1∶3，1∶1混合培养12 d时细胞吸光度值达高峰，且按1∶1混合培养的吸光度值最高；而单纯血管内皮细胞培养10 d吸光度值达高峰；随后各培养方式下细胞吸光度值逐渐下降。提示血管内皮细胞与脂肪干细胞体外联合培养，细胞能够相互促进增殖，1∶1混合比例条件下细胞增殖能力最强

关键词: 增殖, 联合培养, 体外, 血管内皮细胞, 脂肪干细胞, 干细胞

Abstract:

BACKGROUND: Vascular endothelial cells (VECs) have the ability to enhance adipose tissue-derived stem cells (ADSCs) to ossify and new blood vessel can provide nutrition for the ossification of stem cells.
OBJECTIVE: To explore the influence of coculture of VECs and ADSCs on cell proliferation in vitro.
METHODS: We set simple VECs group, 3: 1, 1: 3, 1: 1 coculture group and simple ADSCs group. Cells were incubated in L-DMEM containing fetal bovine serum at 37 ℃ in a 5% CO2 saturated humidity incubator. Morphological changes were observed by inverted microscope. Cell growth curve was drawn using modified MTT assay.
RESULTS AND CONCLUSION: ADSCs and VECs were mixed at 3: 1 and 1: 1 for 14 days. Many synaptic linkages were visible among cells. Partial cells fused into a mass. Cell masses were not observed by other culture methods. With prolonged time, the absorbance of every group was increased gradually and reached a peak in ADSCs, 3: 1, 1: 3 and 1: 1 ratio groups on day 12 and the highest was the 1:1 group, but the time of reaching the peak was on day 10 in the VECs group. The absorbance was gradually reduced under other culture methods. Results have indicated that VECs and ADSCs can stimulate proliferation each other in the system of co-culture in vitro; it can reach the best result when the proportion is 1: 1.

中图分类号: