Department of Biochemistry, University of Colorado, Boulder, CO 80309, USA.
Tissue Eng Part A. 2010 May;16(5):1607-19. doi: 10.1089/ten.TEA.2009.0372.
Fibrin gels are attractive scaffolds useful for neural tissue engineering applications. The objective of this work was to investigate the apoptotic activity, survival, proliferation, and differentiation of a mixed population of primary neural cells composed of neurons and multipotent precursor cells when cultured in fibrin gels prepared with varying concentrations of fibrinogen (5-25 mg/mL fibrinogen) and thrombin (1-125 U/mL thrombin). Within all fibrin gel formulations tested, the level of apoptosis on day 1 was low and cell survival was equivalent to levels in monolayer culture (67%). Proliferation in gels made from 5 to 12.5 mg/mL fibrinogen was also similar to that observed in monolayer culture, though a lower proliferative response was observed in 25 mg/mL fibrinogen formulations. Relative to monolayer culture, cholinergic and dopaminergic neuronal presence was enhanced, whereas glial cell growth was reduced in fibrin gel cultures. The extent to which levels were altered depended on fibrinogen and thrombin concentration. The findings here suggest the importance of fibrinogen and thrombin concentration in differentially regulating the growth and composition of neural cell populations and are of importance for neural tissue engineering strategies focused on the development of implantable scaffolds for treating neurodegenerative disorders.
纤维蛋白凝胶是一种有吸引力的支架,可用于神经组织工程应用。本研究的目的是研究在不同浓度纤维蛋白原(5-25 mg/mL 纤维蛋白原)和凝血酶(1-125 U/mL 凝血酶)制备的纤维蛋白凝胶中培养的由神经元和多能前体细胞组成的混合原代神经细胞群体的凋亡活性、存活、增殖和分化。在所有测试的纤维蛋白凝胶配方中,第 1 天的细胞凋亡水平较低,细胞存活率与单层培养中的水平相当(67%)。在 5 至 12.5 mg/mL 纤维蛋白原制成的凝胶中,增殖也类似于在单层培养中观察到的增殖,尽管在 25 mg/mL 纤维蛋白原制剂中观察到较低的增殖反应。与单层培养相比,纤维蛋白凝胶培养中胆碱能和多巴胺能神经元的存在增强,而神经胶质细胞的生长减少。改变的程度取决于纤维蛋白原和凝血酶的浓度。这些发现表明纤维蛋白原和凝血酶浓度在调节神经细胞群体的生长和组成方面具有重要作用,对于专注于开发用于治疗神经退行性疾病的可植入支架的神经组织工程策略具有重要意义。