Greschat Susanne, Schira Jessica, Küry Patrick, Rosenbaum Claudia, de Souza Silva Maria Angelica, Kögler Gesine, Wernet Peter, Müller Hans Werner
Molecular Neurobiology Laboratory, Heinrich-Heine University, D-40225 Düsseldorf, Germany.
Stem Cells Dev. 2008 Apr;17(2):221-32. doi: 10.1089/scd.2007.0118.
Recently, it has been shown that human unrestricted somatic stem cells (USSCs) from umbilical cord blood represent pluripotent, neonatal, nonhematopoietic stem cells with the potential to differentiate into the neural lineage. However, molecular and functional characterization of the neural phenotype and evaluation of the degree of maturity of the resulting cells are still lacking. In this study, we addressed the question of neuronal differentiation and maturation induced by a defined composition of growth and differentiation factors (XXL medium). We demonstrated the expression of different neuronal markers and their enrichment in USSC cultures during XXL medium incubation. Furthermore, we showed enrichment of USSCs expressing tyrosine hydroxylase (TH), an enzyme specific for dopaminergic neurons and other catecholamine-producing neurons, accompanied by induction of Nurr1, a factor regulating dopaminergic neurogenesis. The functionality of USSCs has been analyzed by patch-clamp recordings and high-performance liquid chromatography (HPLC). Voltage-gated sodium-channels could be identified in laminin-predifferentiated USSCs. In addition, HPLC analysis revealed synthesis and release of the neurotransmitter dopamine by USSC-derived cells, thus correlating well with the detection of TH transcripts and protein. This study provides novel insight into the potential of unrestricted somatic stem cells from human umbilical cord blood to acquire a neuronal phenotype and function.
最近的研究表明，来自脐带血的人类无限制体细胞干细胞（USSCs）代表多能性、新生、非造血干细胞，具有分化为神经谱系的潜力。然而，目前仍缺乏对神经表型的分子和功能特征以及所产生细胞成熟度的评估。在本研究中，我们探讨了由特定组成的生长和分化因子（XXL培养基）诱导的神经元分化和成熟问题。我们证明了在XXL培养基孵育期间，不同神经元标志物的表达及其在USSC培养物中的富集。此外，我们发现表达酪氨酸羟化酶（TH）的USSCs富集，TH是一种对多巴胺能神经元和其他产生儿茶酚胺的神经元特异的酶，同时伴有调节多巴胺能神经发生的因子Nurr1的诱导。通过膜片钳记录和高效液相色谱（HPLC）分析了USSCs的功能。在层粘连蛋白预分化的USSCs中可以鉴定出电压门控钠通道。此外，HPLC分析显示USSC衍生细胞合成并释放神经递质多巴胺，这与TH转录本和蛋白质的检测结果高度相关。本研究为人类脐带血无限制体细胞干细胞获得神经元表型和功能的潜力提供了新的见解。
