Jarmalavičiūtė Akvilė, Tunaitis Virginijus, Strainienė Eglė, Aldonytė Rūta, Ramanavičius Arūnas, Venalis Algirdas, Magnusson Karl-Eric, Pivoriūnas Augustas
Department of Stem Cell Biology, State Research Institute Centre for Innovative Medicine, Žygimantų 9, 01102, Vilnius, Lithuania.
J Mol Neurosci. 2013 Jun 26. doi: 10.1007/s12031-013-0046-0.
Stem cells isolated from human adult tissues represent a promising source for neural differentiation studies in vitro. We have isolated and characterized stem cells from human exfoliated deciduous teeth (SHEDs). These originate from the neural crest and therefore particularly suitable for induction of neural differentiation. We here established a novel three-stage protocol for neural differentiation of SHEDs cells. After adaptation to a serum-free and neurogenic environment, SHEDs were induced to differentiate. This resulted in the formation of stellate or bipolar round-shaped neuron-like cells with subpopulations expressing markers of sensory neurons (Brn3a, peripherin) and glia (myelin basic protein). Commercial PCR array analyses addressed the expression profiles of genes related to neurogenesis and cAMP/calcium signalling. We found distinct evidence for the upregulation of genes regulating the specification of sensory (MAF), sympathetic (midkine, pleitrophin) and dopaminergic (tyrosine hydroxylase, Nurr1) neurons and the differentiation and support of myelinating and non-myelinating Schwann cells (Krox24, Krox20, apolipoprotein E). Moreover, for genes controlling major developmental signalling pathways, there was upregulation of BMP (TGF β-3, BMP2) and Notch (Notch 2, DLL1, HES1, HEY1, HEY2) in the differentiating SHEDs. SHEDs treated according to our new differentiation protocol gave rise to mixed neuronal/glial cell cultures, which opens new possibilities for in vitro studies of neuronal and glial specification and broadens the potential for the employment of such cells in experimental models and future treatment strategies.
从成人组织中分离出的干细胞是体外神经分化研究的一个有前景的来源。我们已经从人脱落乳牙（SHEDs）中分离并鉴定了干细胞。这些干细胞起源于神经嵴，因此特别适合诱导神经分化。我们在此建立了一种新的用于SHEDs细胞神经分化的三阶段方案。在适应无血清和神经源性环境后，诱导SHEDs分化。这导致形成星状或双极圆形的神经元样细胞，其中亚群表达感觉神经元（Brn3a、外周蛋白）和神经胶质细胞（髓磷脂碱性蛋白）的标志物。商业PCR阵列分析研究了与神经发生和cAMP/钙信号相关的基因表达谱。我们发现了明确的证据表明，调节感觉神经元（MAF）、交感神经元（中期因子、多效生长因子）和多巴胺能神经元（酪氨酸羟化酶、Nurr1）特异性的基因以及有髓和无髓雪旺细胞（Krox24、Krox20、载脂蛋白E）的分化和支持相关基因上调。此外，对于控制主要发育信号通路的基因，在分化的SHEDs中BMP（TGFβ-3、BMP2）和Notch（Notch 2、DLL1、HES1、HEY1、HEY2）上调。按照我们新的分化方案处理的SHEDs产生了混合的神经元/神经胶质细胞培养物，这为神经元和神经胶质特异性的体外研究开辟了新的可能性，并拓宽了在实验模型和未来治疗策略中使用此类细胞的潜力。
