细胞外基质蛋白模拟物对绒毛膜来源间充质干细胞黏附与增殖的影响
Effects of ECM protein mimetics on adhesion and proliferation of chorion derived mesenchymal stem cells.
作者信息
Kim Ji-Hyun, Jekarl Dong Wook, Kim Myungshin, Oh Eun-Jee, Kim Yonggoo, Park In Yang, Shin Jong Chul
机构信息
1. Department of Laboratory Medicine College of Medicine, The Catholic University of Korea, Seoul, South Korea; ; 3. Department of Biomedical Science, College of Medicine, The Catholic University of Korea, Seoul, South Korea.
1. Department of Laboratory Medicine College of Medicine, The Catholic University of Korea, Seoul, South Korea;
出版信息
Int J Med Sci. 2014 Feb 6;11(3):298-308. doi: 10.7150/ijms.6672. eCollection 2014.
BACKGROUND
We evaluated the effects of fibronectin, collagen, cadherin, and laminin based extracellular matrix (ECM) protein mimetics coated with mussel derived adhesive protein (MAP) on adhesion and proliferation of chorionic mesenchymal stem cells (cMSCs).
METHODS
Human placental chorionic tissues from term third-trimester pregnancies (n=3) were used. The cMSCs were cultured on rationally designed ECM protein mimetics coated with MAP on plastic surfaces with the addition of reduced fetal bovine serum (0.5%, 1% FBS). Adhesion capabilities were monitored by a real time cell analysis system (RTCA) utilizing an impedance method. Proliferation capabilities were monitored by RTCA and MTS assay.
RESULTS
Of the ECM protein mimetics tested, GRGDSP(FN) coated surfaces exhibited the highest adhesion and proliferation capabilities on RTCA at FBS concentration of 0.5% and 1%. When 0.5% FBS was added to ECM protein mimetics during the MTS assay, GRGDSP(FN), REDV(FN), and collagen mimetics, GPKGAAGEPGKP(ColI) showed higher cMSCs proliferation compared with the control. When 1% FBS was added, GRGDSP(FN) and TAIPSCPEGTVPLYS(ColIV) showed significant cMSCs proliferation capacity.
CONCLUSIONS
Fibronectin mimetics, GRGDSP(FN) amino acid sequence showed the highest adhesion and proliferation capabilities. In addition, results from RTCA assessment of cell viability correlated well with the tetrazolium-based MTS assay.
背景
我们评估了涂有贻贝衍生黏附蛋白（MAP）的基于纤连蛋白、胶原蛋白、钙黏蛋白和层粘连蛋白的细胞外基质（ECM）蛋白模拟物对绒毛膜间充质干细胞（cMSCs）黏附和增殖的影响。
方法
使用来自足月妊娠晚期（n = 3）的人胎盘绒毛膜组织。将cMSCs培养在涂有MAP的合理设计的ECM蛋白模拟物上，置于塑料表面，并添加减少的胎牛血清（0.5%，1% FBS）。通过利用阻抗法的实时细胞分析系统（RTCA）监测黏附能力。通过RTCA和MTS测定法监测增殖能力。
结果
在所测试的ECM蛋白模拟物中，涂有GRGDSP（FN）的表面在FBS浓度为0.5%和1%时，在RTCA上表现出最高的黏附和增殖能力。在MTS测定期间向ECM蛋白模拟物中添加0.5% FBS时，GRGDSP（FN）、REDV（FN）和胶原蛋白模拟物GPKGAAGEPGKP（ColI）与对照组相比显示出更高的cMSCs增殖。当添加1% FBS时，GRGDSP（FN）和TAIPSCPEGTVPLYS（ColIV）显示出显著的cMSCs增殖能力。
结论
纤连蛋白模拟物GRGDSP（FN）氨基酸序列表现出最高的黏附和增殖能力。此外，RTCA细胞活力评估结果与基于四唑盐的MTS测定法相关性良好。
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