Seyedhassantehrani Negar, Li Yongchao, Yao Li
Department of Biological Sciences, Wichita State University, Fairmount 1845, Wichita, KS 67260, USA.
Integr Biol (Camb). 2016 May 16;8(5):624-34. doi: 10.1039/c6ib00003g. Epub 2016 Apr 15.
Astrocytes play a critical role in supporting the normal physiological function of neurons in the central nervous system (CNS). Astrocyte transplantation can potentially promote axonal regeneration and functional recovery after spinal cord injury (SCI). Fibrin and collagen hydrogels provide growth-permissive substrates and serve as carriers for therapeutic cell transplantation into an injured spinal cord. However, the application of fibrin and collagen hydrogels may be limited due to their relatively rapid degradation rate in vivo. In this study, immature astrocytes isolated from neonatal rats were grown in fibrin hydrogels containing aprotinin and collagen hydrogels crosslinked with poly(ethylene glycol) ether tetrasuccinimidyl glutarate (4S-StarPEG), and the cell behavior in these hydrogels was studied. The cell viability of astrocytes in the hydrogels was tested using the LIVE/DEAD® assay and the AlamarBlue® assay, and this study showed that astrocytes maintained good viability in these hydrogels. The cell migration study showed that astrocytes migrated in the fibrin and collagen hydrogels, and the migration speed is similar in these hydrogels. The crosslinking of collagen hydrogels with 4S-StarPEG did not change the astrocyte migration speed. However, the addition of aprotinin in the fibrin hydrogel inhibited astrocyte migration. The expression of chondroitin sulfate proteoglycan (CSPG), including NG2, neurocan, and versican, by astrocytes grown in the hydrogels was analyzed by quantitative RT-PCR. The expression of NG2, neurocan, and versican by the cells in these hydrogels was not significantly different.
星形胶质细胞在支持中枢神经系统(CNS)中神经元的正常生理功能方面发挥着关键作用。星形胶质细胞移植可能会促进脊髓损伤(SCI)后的轴突再生和功能恢复。纤维蛋白和胶原蛋白水凝胶提供有利于生长的底物,并作为治疗性细胞移植到受损脊髓中的载体。然而,由于纤维蛋白和胶原蛋白水凝胶在体内的降解速度相对较快,其应用可能会受到限制。在本研究中,从新生大鼠分离的未成熟星形胶质细胞在含有抑肽酶的纤维蛋白水凝胶和与聚(乙二醇)醚四琥珀酰亚胺戊二酸酯(4S-StarPEG)交联的胶原蛋白水凝胶中生长,并研究了这些水凝胶中的细胞行为。使用LIVE/DEAD®检测法和AlamarBlue®检测法测试了水凝胶中星形胶质细胞的细胞活力,该研究表明星形胶质细胞在这些水凝胶中保持良好的活力。细胞迁移研究表明,星形胶质细胞在纤维蛋白和胶原蛋白水凝胶中迁移,并且在这些水凝胶中的迁移速度相似。胶原蛋白水凝胶与4S-StarPEG的交联并没有改变星形胶质细胞的迁移速度。然而,在纤维蛋白水凝胶中添加抑肽酶会抑制星形胶质细胞的迁移。通过定量RT-PCR分析了在水凝胶中生长的星形胶质细胞硫酸软骨素蛋白聚糖(CSPG)的表达,包括NG2、神经黏蛋白和多功能蛋白聚糖。这些水凝胶中细胞的NG2、神经黏蛋白和多功能蛋白聚糖的表达没有显著差异。