Völlner Florian, Ernst Wolfgang, Driemel Oliver, Morsczeck Christian
Institute of Human Genetics, Franz-Josef Strauss Allee 11, University of Regensburg, 93053 Regensburg, Germany.
Differentiation. 2009 Jun;77(5):433-41. doi: 10.1016/j.diff.2009.03.002. Epub 2009 Apr 25.
Human dental follicle cells (DFCs) derived from wisdom teeth are precursor cells for cementoblasts. In this study, we recognized that naïve DFCs express constitutively the early neural cell marker beta-III-tubulin. Interestingly, DFCs formed beta-III-tubulin-positive neurosphere-like cell clusters (NLCCs) on low-attachment cell culture dishes in serum-replacement medium (SRM). For a detailed examination of the neural differentiation potential, DFCs were cultivated in different compositions of SRM containing supplements such as N2, B27, G5 and the neural stem cell supplement. Moreover, these cell culture media were combined with different cell culture substrates such as gelatin, laminin, poly-L-ornithine or poly-L-lysine. After cultivation in SRM, DFCs differentiated into cells with small cell bodies and long cellular extrusions. The expression of nestin, beta-III-tubulin, neuron-specific enolase (NSE) and neurofilament was up-regulated in SRM supplemented with G5, a cell culture supplement for glial cells, and the neural stem cell supplement. DFCs formed NLCCs and demonstrated an increased gene expression of neural cell markers beta-III-tubulin, NSE, nestin and for small neuron markers such as neuropeptides galanin (GAL) and tachykinin (TAC1) after cultivation on poly-L-lysine. For a further neural differentiation NLCC-derived cells were sub-cultivated on laminin and poly-L-ornithine cell culture substrate. After 2 weeks of differentiation, DFCs exposed neural-like cell morphology with small neurite-like cell extrusions. These cells differentially express neurofilament and NSE, but only low levels of beta-III-tubulin and nestin. In conclusion, we demonstrated the differentiation of human DFCs into neuron-like cells after a two-step strategy for neuronal differentiation.
源自智齿的人牙囊细胞（DFCs）是成牙骨质细胞的前体细胞。在本研究中，我们发现未分化的DFCs组成性表达早期神经细胞标志物β-III-微管蛋白。有趣的是，DFCs在血清替代培养基（SRM）中的低附着细胞培养皿上形成了β-III-微管蛋白阳性的神经球样细胞簇（NLCCs）。为了详细研究神经分化潜能，将DFCs培养在含有N2、B27、G5等补充剂以及神经干细胞补充剂的不同组成的SRM中。此外，这些细胞培养基还与不同的细胞培养底物如明胶、层粘连蛋白、聚-L-鸟氨酸或聚-L-赖氨酸结合使用。在SRM中培养后，DFCs分化为具有小细胞体和长细胞突起的细胞。在添加了G5（一种用于神经胶质细胞的细胞培养补充剂）和神经干细胞补充剂的SRM中，巢蛋白、β-III-微管蛋白、神经元特异性烯醇化酶（NSE）和神经丝的表达上调。DFCs形成了NLCCs，并且在聚-L-赖氨酸上培养后，神经细胞标志物β-III-微管蛋白、NSE、巢蛋白以及小神经元标志物如神经肽甘丙肽（GAL）和速激肽（TAC1）的基因表达增加。为了进一步进行神经分化，将NLCC衍生的细胞在层粘连蛋白和聚-L-鸟氨酸细胞培养底物上进行传代培养。分化2周后，DFCs呈现出具有小神经突样细胞突起的神经样细胞形态。这些细胞差异表达神经丝和NSE，但仅低水平表达β-III-微管蛋白和巢蛋白。总之，我们通过两步神经元分化策略证明了人DFCs可分化为神经元样细胞。
