Janowski Miroslaw, Lukomska Barbara, Domanska-Janik Krystyna
Department of NeuroRepair, Mossakowski Medical Research Centre, Polish Academy of Sciences, Warsaw, Poland.
Acta Neurobiol Exp (Wars). 2011;71(1):24-35. doi: 10.55782/ane-2011-1820.
Many types of neural progenitors from various sources have been evaluated for therapy of CNS disorders. Prerequisite for success in cell therapy is the ability for transplanted cells to reach appropriate target such as stroke lesion. We have established neural stem cell line from human umbilical cord blood neural stem (HUCB-NSC). In the present study we evaluated migratory capabilities of cells (HUCB-NSC) and the presence of various migration-related receptors. Immunocytochemical analysis revealed abundant expression of CXCR4, PDGFR-alpha, PDGFR-beta, c-Met, VEGFR, IGF-1R and PSA-NCAM receptors in non-adherent population of HUCB-NSC cultured in serum free (SF) conditions (SF cells). Biological activity of selected receptors was confirmed by HUCB-NSC in vitro migration towards SDF-1 and IGF-1 ligands. Additionally, rat brain-derived homogenates have been assessed for their chemoattractive activity of HUCB-NSC. Our experiments unveiled that brain tissue was more attracted for HUCB-NSC than single ligands with higher potency of injured than intact brain. Moreover, adherent HUCB-NSC cultured in low serum (LS) conditions (LS cells) were employed to investigate an impact of different extracellular matrix (ECM) proteins on cell motility. It turned out that laminin provided most permissive microenvironment for cell migration, followed by fibronectin and gelatin. Unexpected nuclear localization of CXCR4 in SF cells prompted us to characterize intracellular pattern of this expression in relation to developmental stage of cells cultured in different conditions. Continuous culture of LS cells revealed cytoplasmatic pattern of CXCR4 expression while HUCB-NSC cultured in high serum conditions (HS cells) resulted in gradual translocation of CXCR4 from nucleus to cytoplasm and then to arising processes. Terminal differentiation of HUCB-NSC was followed by CXCR4 expression decline.
多种来源的多种类型神经祖细胞已被评估用于中枢神经系统疾病的治疗。细胞治疗成功的前提是移植细胞能够到达合适的靶点，如中风病灶。我们已从人脐带血神经干细胞（HUCB-NSC）建立了神经干细胞系。在本研究中，我们评估了细胞（HUCB-NSC）的迁移能力以及各种与迁移相关受体的存在情况。免疫细胞化学分析显示，在无血清（SF）条件下培养的HUCB-NSC非贴壁群体（SF细胞）中，CXCR4、血小板衍生生长因子受体α（PDGFR-α）、血小板衍生生长因子受体β（PDGFR-β）、c-Met、血管内皮生长因子受体（VEGFR）、胰岛素样生长因子-1受体（IGF-1R）和多唾液酸神经细胞黏附分子（PSA-NCAM）受体表达丰富。通过HUCB-NSC体外向基质细胞衍生因子-1（SDF-1）和胰岛素样生长因子-1（IGF-1）配体迁移，证实了所选受体的生物学活性。此外，还评估了大鼠脑源性匀浆对HUCB-NSC的趋化活性。我们的实验表明，与单个配体相比，脑组织对HUCB-NSC的吸引力更大，损伤脑组织的趋化活性比完整脑组织更强。此外，采用在低血清（LS）条件下培养的贴壁HUCB-NSC（LS细胞）来研究不同细胞外基质（ECM）蛋白对细胞运动性的影响。结果表明，层粘连蛋白为细胞迁移提供了最宽松的微环境，其次是纤连蛋白和明胶。SF细胞中CXCR4意外的核定位促使我们表征这种表达的细胞内模式与在不同条件下培养的细胞发育阶段的关系。LS细胞的连续培养显示CXCR4表达呈细胞质模式，而在高血清条件下培养的HUCB-NSC（HS细胞）导致CXCR4从细胞核逐渐转移到细胞质，然后转移到新生的突起中。HUCB-NSC的终末分化伴随着CXCR4表达下降。
