纤维蛋白胶对骨髓源性心肌干细胞增殖、迁移和缺血缺氧后的保护作用

目的探讨纤维蛋白胶（FG）对骨髓源性心肌干细胞（MCSC）增殖、迁移和存活的作用,为临床应用干细胞移植治疗心肌梗死等缺血性疾病提供实验手段和理论依据。方法将MCSC接种于FG中制备FG-MCSC,经AO/EB染色选择合适浓度的FG,用细胞计数法检测MCSC在FG中的增殖,通过损伤和自然迁移实验评价MCSC从FG中的迁出。建立细胞缺血缺氧模型（OGD）,用AO/EB染色和乳酸脱氢酶（LDH）泄漏法检测FG对MCSC的保护作用。结果 MCSC在低浓度FG中保持较高的活力和增殖能力。接种后1d和2d,仅有少量细胞从FG的刮除线边缘迁出。在接种后3d,有大量细胞从FG中迁出。经缺血缺氧处理后,MCSCOGD组中存活、凋亡和坏死细胞数目分别为14.09±2.05、61.11±5.20和24.80±4.99。与MCSC OGD组相比,FG-MCSC OGD组MCSC的存活数目（61.01±8.86）较高,凋亡（31.67±8.27）和坏死（7.33±2.70）细胞数目较低。MCSC经缺血缺氧处理后,FG-MCSC OGD组LDH的水平明显低于MCSC OGD组。结论低浓度FG作为一种天然生物材料可以为MC-SC的生长提供良好的微环境。在缺血缺氧条件下,低浓度FG能够保护MCSC免受缺血缺氧损伤,促进细胞存活。

Effects of Fibrin Glue on Proliferation, Migration and Ischemic Hypoxia Protection of Marrow - derived Cardiac Stem Cell

Objective To investigate effects of fibrin glue（FG） on the proliferation,migration and survival of marrow -derived cardiac stem cell （MCSC）,and to provide experimental methods and theoretical basis for clinical application of stem cell transplantation in the treatment of ischemic diseases,such as myocardial infarction. Methods MCSC were inoculated in FG to prepare the FG- MCSC. After selecting the appropriate concentration of FG by AO/ EB staining, MCSC proliferation was detected by cell counting. Migration of MCSC from the FG was evaluated through the damage and natural migration assay. The protective effect of FG on MCSC was ex- amined with AO/EB staining and lactate dehydrogenase （LDH） leakage assay after the establishment of cellular hypoxia- ischemia （OGD） model Results The MCSC maintained a high viability and proliferation activity in the FG of low concen tration. At 1 d and 2 d after seeding,only a few cells moved out from the edge of the scrape line of the FG. at 3 days after seeding. There were a large number of cells moved out from the FG. In the Mcsc group,the number of survival,apoptosis and necrosis cells were 14.09±2.05,61.11± 5.20 and 24.80±4.99 after ischemia and hypoxia treatment. Compared with the MCSC OGD group,number of survived cells （61.01± 8.86） was higher,and number of apoptosis ceils （31.67±8.27） and necrosis cells （7.33±2.70） was lower in the group of FG - MCSC OGD The LDH level in the group of FG - MCSC OGD was significantly lower than that in MCSC OGD group after isehemia and hypoxia treatment. Conclusion FG of low concentration,as a natural biological material,can provide favorable microenvironment for the growth of MCSC. In the ischemia and hypoxia conditions,FG of low concentration may protect MCSC from injury by ischemia and hypoxia,and promote cell survival.