Gao Songtao, Zheng Yan, Cai Qiqing, Wu Xuejian, Yao Weitao, Wang Jiaqiang
Department of Orthopedics, The Affiliated Tumor Hospital of Zhengzhou University, Zhengzhou, China.
Department of Radiology, The First Affiliated Hospital of Zhengzhou University, Zhengzhou, China.
Arch Med Sci. 2015 Aug 12;11(4):886-92. doi: 10.5114/aoms.2015.53310. Epub 2015 Aug 11.
The aim of the study was to explore an effective method to induce adipose-derived stem cells (ADSCs) to differentiate into Schwann-like cells in vitro.
Reagents were applied in two different ways (Dezawa inducing method and modified inducing method) in which inducers including β-mercaptoethanol (β-ME), all-trans-retinoic acid (ATRA), type I collagenase, forskolin, heregulin, basic fibroblast growth factor (BFGF) and brain-derived neurotrophic factor (BDNF) were used in different ways to induce ADSCs of rats to differentiate into Schwann-like cells. After induction, the cell morphologic characteristics and the cellular immunohistochemical staining positive rate of anti-S100 and anti-GFAP (glial fibrillary acidic protein) antibodies and the gray value of immunocytochemical dye with anti-S100 and anti-GFAP antibodies and cell activity measured by the MTT method were compared with each other to evaluate the induction effects.
Both methods can induce differentiation of ADSCs of rats into Schwann-like cells, but the cellular morphology of the modified method was more similar to Schwann cells than that of the Dezawa inducing method, there was a higher cellular immunohistochemical staining positive rate and staining grey value in immunocytochemical dye with anti-S100 and anti-GFAP antibodies, and less damage in the cell activity of the modified inducing method than that of the Dezawa inducing method.
The effect of the modified method to induce ADSCs to differentiate into Schwann-like cells in vitro is superior to that of the Dezawa inducing method.
本研究的目的是探索一种在体外诱导脂肪来源干细胞(ADSCs)分化为雪旺氏样细胞的有效方法。
试剂采用两种不同的方式(Dezawa诱导法和改良诱导法)应用,其中诱导剂包括β-巯基乙醇(β-ME)、全反式维甲酸(ATRA)、I型胶原酶、福司可林、神经调节蛋白、碱性成纤维细胞生长因子(BFGF)和脑源性神经营养因子(BDNF),以不同方式用于诱导大鼠ADSCs分化为雪旺氏样细胞。诱导后,比较细胞形态学特征、抗S100和抗胶质纤维酸性蛋白(GFAP)抗体的细胞免疫组化染色阳性率、抗S100和抗GFAP抗体免疫细胞化学染色的灰度值以及通过MTT法测定的细胞活性,以评估诱导效果。
两种方法均可诱导大鼠ADSCs分化为雪旺氏样细胞,但改良法的细胞形态比Dezawa诱导法更类似于雪旺细胞,抗S100和抗GFAP抗体免疫细胞化学染色的细胞免疫组化染色阳性率和染色灰度值更高,改良诱导法的细胞活性损伤比Dezawa诱导法更小。
改良法在体外诱导ADSCs分化为雪旺氏样细胞的效果优于Dezawa诱导法。