Institute of Bioscience, University Putra Malaysia, Kuala Lumpur, Malaysia.
Cytotechnology. 2015 May;67(3):449-60. doi: 10.1007/s10616-014-9703-6. Epub 2014 Oct 26.
Many neural disorders are characterized by the loss of one or several types of neural cells. Human umbilical cord-derived mesenchymal cells (hUCMs) are capable of differentiating into neuron, astroglia-like and oligodendrocyte cell types. However, a reliable means of inducing the selective differentiation of hUCMs into neural cells in vitro has not yet been established. For induction of neural differentiation, hUCMs were seeded onto sterile glass slides and six various cocktails using a base medium (DMEM/LG) supplemented with 10 % FBS, retinoic acid (RA), dimethyl sulfoxide (DMSO), epidermal growth factor (EGF) and fibroblast growth factor (FGF) were used to compare their effect on neuronal, astrocyte and oligodandrocyte differentiation. The hUCMs were positive for mesenchymal markers, while they were negative for hematopoietic markers. Differentiation to adipogenic and osteogenic lineage was detected in these cells. Our data revealed that the cocktail consisting of DMEM/LG, FBS, RA, FGF, and EGF (DF/R/Fg/E group) induced hUCM cells to express the highest percentage of nestin, ß-tubulin III, neurofilament, and CNPase. The DF/Ds/Fg/E group led to the highest percentage of GFAP expression. While the expression levels of NF, GFAP, and CNPase were the lowest in the DF group. The least percentage of nestin and ß-tubulin III expression was observed in the DF/Ds group. We may conclude that FGF and EGF are important inducers for differentiation of hUCMs into neuron, astrocyte and oligodendrocyte. RA can induce hUCMs to differentiate into neuron and oligodendrocyte while for astrocyte differentiation DMSO had a pivotal role.
许多神经紊乱疾病的特征是一种或多种类型的神经细胞丧失。人脐带间充质细胞(hUCMs)能够分化为神经元、星形胶质样细胞和少突胶质细胞。然而,尚未建立一种可靠的方法来诱导 hUCMs 在体外选择性分化为神经细胞。为了诱导神经分化,将 hUCMs 接种到无菌玻璃幻灯片上,使用含有 10% FBS、维甲酸(RA)、二甲亚砜(DMSO)、表皮生长因子(EGF)和纤维生长因子(FGF)的基础培养基(DMEM/LG)的六种不同的鸡尾酒来比较它们对神经元、星形胶质细胞和少突胶质细胞分化的影响。hUCMs 对间充质标志物呈阳性,而对造血标志物呈阴性。在这些细胞中检测到向脂肪和成骨谱系的分化。我们的数据显示,由 DMEM/LG、FBS、RA、FGF 和 EGF 组成的鸡尾酒(DF/R/Fg/E 组)诱导 hUCM 细胞表达最高百分比的巢蛋白、β-微管蛋白 III、神经丝和 CNPase。DF/Ds/Fg/E 组导致 GFAP 表达的百分比最高。而 NF、GFAP 和 CNPase 的表达水平在 DF 组中最低。DF/Ds 组中观察到巢蛋白和β-微管蛋白 III 的表达百分比最低。我们可以得出结论,FGF 和 EGF 是诱导 hUCM 分化为神经元、星形胶质细胞和少突胶质细胞的重要诱导剂。RA 可以诱导 hUCM 分化为神经元和少突胶质细胞,而 DMSO 在星形胶质细胞分化中起着关键作用。