Safford K M, Safford S D, Gimble J M, Shetty A K, Rice H E
Department of Surgery, Duke University Medical Center, Durham, NC 27710, USA.
Exp Neurol. 2004 Jun;187(2):319-28. doi: 10.1016/j.expneurol.2004.01.027.
Neural tissue has limited capacity for intrinsic repair after injury, and the identification of alternate sources of neuronal stem cells has broad clinical potential. Preliminary studies have demonstrated that adipose-derived adult stromal (ADAS) cells are capable of differentiating into mesenchymal and non-mesenchymal cells in vitro, including cells with select characteristics of neuronal/glial tissue. In this study, we extended these observations to test the hypothesis that murine (mu) ADAS cells can be induced to exhibit characteristics of neuronal and glial tissue by exposure to a cocktail of induction agents. We characterized the differentiation of muADAS cells in vitro using immunohistochemistry and immunoblotting, and examined whether these cells respond to the glutamate agonist N-methyl-D-aspartate (NMDA). We found that induced muADAS cells express proteins indicative of neuronal/glial cells, including nestin, GFAP, S-100, NeuN, MAP2, tau, and beta-III tubulin. Induced muADAS cells express gamma-aminobutyric acid (GABA), the NR-1 and NR-2 subunits of the glutamate receptor, GAP-43, synapsin I, and voltage-gated calcium channels. Finally, induced muADAS cells demonstrate decreased viability in response to NMDA. These findings suggest that muADAS cells can be induced to exhibit several phenotypic, morphologic, and excitotoxic characteristics consistent with developing neuronal and glial tissue.
神经组织损伤后自身修复能力有限,而寻找替代的神经干细胞来源具有广泛的临床潜力。初步研究表明,脂肪来源的成人基质(ADAS)细胞在体外能够分化为间充质和非间充质细胞,包括具有某些神经/胶质组织特征的细胞。在本研究中,我们拓展了这些观察结果,以检验如下假设:通过暴露于一组诱导剂,小鼠(mu)ADAS细胞能够被诱导表现出神经和胶质组织的特征。我们使用免疫组织化学和免疫印迹法在体外对muADAS细胞的分化进行了表征,并检测这些细胞是否对谷氨酸激动剂N-甲基-D-天冬氨酸(NMDA)产生反应。我们发现,诱导后的muADAS细胞表达指示神经/胶质细胞的蛋白质,包括巢蛋白、胶质纤维酸性蛋白(GFAP)、S-100、神经元核抗原(NeuN)、微管相关蛋白2(MAP2)、微管蛋白(tau)和β-III微管蛋白。诱导后的muADAS细胞表达γ-氨基丁酸(GABA)、谷氨酸受体的NR-1和NR-2亚基、生长相关蛋白43(GAP-43)、突触素I和电压门控钙通道。最后,诱导后的muADAS细胞对NMDA产生反应后活力下降。这些发现表明,muADAS细胞能够被诱导表现出与正在发育的神经和胶质组织一致的几种表型、形态学和兴奋毒性特征。