Farrokhfar Samaneh, Tiraihi Taki, Movahedin Mansoureh, Azizi Hossein
Department of Anatomy, Faculty of Medical Sciences, Tarbiat Modares University, Tehran, Iran.
Department of Anatomy, Faculty of Medical Sciences, Tarbiat Modares University, P.O.BOX.14115-331 Tehran, Iran.
Iran J Basic Med Sci. 2020 Apr;23(4):544-550. doi: 10.22038/ijbms.2020.41582.9820.
Adipose-derived stem cells (ADSCs), with suitable and easy access, are multipotential cells that have the ability for differentiation into other mesodermal and transdifferentiate into neural phenotype cells. In this study, Lithium chloride (LiCl) was used for transdifferentiation of rat ADSCs into neuron-like cells (NLCs).
ADSCs were isolated from the rats' perinephric region using Dulbecco΄s Modified Eagle΄s Medium (DMEM) with Fetal Bovine Serum (FBS), cultured for 3 passages, characterized by flowcytometry and differentiation into adipogenic and osteogenic phenotypes. The ADSCs were exposed to 0.1, 0.5, 1, 1.5, 2, 5, and 10 millimolar (mM) LiCl without serum for 24 hr. The optimum dose of LiCl was selected according the maximum viability of cells. The expression of neurofilament light chain (NfL), neurofilament high chain (NfH), and nestin was evaluated by immunocytochemistry. Quantitative reverse transcription polymerase chain reaction (qRT-PCR) was used to evaluate the amount of synaptophysin, neurogenin-1, neuroD1, NfL, NfH, and nestin genes' expression in ADSCs and NLCs.
The optimum dose of LiCl was 1 mM in 24 hr. The transdifferentiated ADSCs showed cytoplasmic extension with synapse-like formation. Synaptophysin, neurogenin-1, neuroD1, NfL, NfH, and nestin genes were significantly expressed more in NLCs than in ADSCs.
LiCl can induce ADSCs into neural phenotype cells with higher expression of neural and neuronal genes.
脂肪来源干细胞（ADSCs）获取便捷，是具有多能性的细胞，能够分化为其他中胚层细胞，并转分化为神经表型细胞。在本研究中，使用氯化锂（LiCl）将大鼠ADSCs转分化为神经元样细胞（NLCs）。
使用含有胎牛血清（FBS）的杜氏改良 Eagle 培养基（DMEM）从大鼠肾周区域分离ADSCs，培养3代，通过流式细胞术进行表征，并诱导其分化为脂肪生成和骨生成表型。将ADSCs在无血清条件下分别暴露于0.1、0.5、1、1.5、2、5和10毫摩尔（mM）的LiCl中24小时。根据细胞的最大活力选择LiCl的最佳剂量。通过免疫细胞化学评估神经丝轻链（NfL）、神经丝重链（NfH）和巢蛋白的表达。采用定量逆转录聚合酶链反应（qRT-PCR）评估ADSCs和NLCs中突触素、神经生成素-1、神经D1、NfL、NfH和巢蛋白基因的表达量。
LiCl的最佳剂量为24小时1 mM。转分化的ADSCs表现出细胞质延伸并形成突触样结构。突触素、神经生成素-1、神经D1、NfL、NfH和巢蛋白基因在NLCs中的表达明显高于ADSCs。
LiCl可诱导ADSCs转分化为神经表型细胞，使其神经和神经元基因表达更高。
