Differentiation of mesenchymal stem cells into myofibroblasts under the induction of tumor microenvironment in rabbits
10.3969/j.issn.1673-8225.2009.45.026
- VernacularTitle:兔骨髓间充质干细胞在肿瘤微环境诱导下向肌纤维母细胞的分化
- Author:
Han LIU
;
Jun CHEN
;
Zhishun XU
- Publication Type:Journal Article
- From:
Chinese Journal of Tissue Engineering Research
2009;13(45):8939-8943
- CountryChina
- Language:Chinese
-
Abstract:
BACKGROUND:Studies have shown that mesenchymsi stem cells (MSCs) participate in tumor stroma remodeling,but the mechanism is not clear.However,myofibroblast is related with tumor strema remodeling.OBJECTIVE:To investigate whether MSCs can differentiate into myofibroblast under the induction of tumor microenvironment.DESIGN,TIME AND SETING:This in vitro observation and in vivo transplantation cytology experiment was performed at institute of Low Temperature Medicine,Qilu Hospital of Shandong University from June 2006 to December 2007.MATERIALS:Twelve male New Zealand white rabbits,with 3-month-old,were purchased from Shandong Academy of Agricultural Sciences.METHODS:MSCs were obtained through bone marrow aspirates in male New Zealand rabbits and cultured by density gradient method.The 2~(nd) passage of MSCs were labeled with DAPI,and adjusted cell density to 5×10~(10) /L.The cells were assigned into experimental and control groups.For the experimental group,the passage two MSCs were cultured in low-glucose DMEM containing 10% fetal bovine serum and 30% VX2-cultured supernatant.For the control group,passage two MSCs were cultured in low-glucose DMEM containing 10% fetal bovine serum.Also,DAPI labeled self passage 2 MSCs of avery animal were delivered uniformly via direct surgical intratumoral injection to observe the differentiation of MSCs in tumor tissue.MAIN OUTCOME MEASURES:Immunofluorescense and RT-PCR were used to test the expression of α-SMA and Vimentin in MSCs of both groups 14 days later.At 3 weeks after VX2.intratumoral injection,Immunofluorescence was used to observe the distribution and differentiation of MSCs in tumor tissue.RESULTS:After stimulated with 30% VX2-cultured supematant for 2 weeks,Cy3 immunofluorescance showed that the expressions of α-SMA and Vimentin were strongly expressed in the experimental group,which was weakly expressed in the control group.Compared to the control group,the levels of α-SMA and Vimentin mRNA were obviously increased in the experimental group (P ＜ 0.01,P ＜ 0.05).At 3 weeks after MSCs intratumoral injection,these MSCs mainly distributed in tumor stroma and the expression of α-SMA and Vimentin in MSCs were also obviously increased,which demonstrated that the MSCs had differentiated into myofibroblasts.CONCLUSIONS:Under the induction of tumor microenivironment,MSCs can differentiate into myofibroblast,accordingly,accelerate the growth of tumor.
